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Fungal Diversity

, Volume 91, Issue 1, pp 5–230 | Cite as

The family Amanitaceae: molecular phylogeny, higher-rank taxonomy and the species in China

  • Yang-Yang Cui
  • Qing Cai
  • Li-Ping Tang
  • Jian-Wei Liu
  • Zhu L. Yang
Article

Abstract

Mushrooms in the basidiomycete family Amanitaceae are very important both economically and ecologically. However, the delimitation of the family is still controversial, in part due to limited taxon sampling and in part because of insufficient gene fragment employed for molecular phylogenetic analyses. Furthermore, species diversity in the family is likely to have been largely underestimated, due to morphological similarity between taxa and phenotypic plasticity. In this study, we examined 1190 collections, including 1008 Chinese and 182 external ones, and performed the first comprehensive phylogenetic analyses of Amanitaceae using multi-locus sequence data. To test the monophyly of the Amanitaceae, a concatenated (nrLSU, rpb1, and rpb2) dataset of 200 taxa of the order Agaricales was analyzed. To infer the phylogeny of Amanitaceae, a concatenated nrLSU, tef1-α, rpb2 and β-tubulin dataset (3010 sequences from ca. 890 samples with 2309 newly generated sequences) was used. In this dataset, 252 sequences from the types of 77 species were provided. Our results indicate that Amanitaceae is a monophyletic group, and consists of five genera, namely Amanita, Catatrama, Limacella, Limacellopsis and Myxoderma. It is clear that Catatrama is closely related to Limacella, however, the phylogenetic relationships among these genera remain largely unresolved. Amanita contains 95% of the species in the family, and is here divided into three subgenera and eleven sections (subgen. Amanita, containing: sect. Amanita, sect. Amarrendiae, sect. Caesareae and sect. Vaginatae; subgen. Amanitina, containing: sect. Amidella, sect. Arenariae, sect. Phalloideae, sect. Roanokenses, sect. Strobiliformes and sect. Validae; and subgen. Lepidella, containing sect. Lepidella). Subgen. Lepidella occupies the basal position in the genus. One-hundred and sixty-two species of Amanitaceae known from China are treated in this study, including 50 novel species and 112 known taxa. Amanita gleocystidiosa, A. pyriformis, A. atrofusca, A. subjunquillea var. alba and A. areolata are treated as synonyms of A. sychnopyramis f. subannulata, A. orientigemmata, A. umbrinolutea, A. subjunquillea and A. zangii, respectively. 26 extralimital taxa including a novel species, namely Catatrama indica, were included in our study to allow us to make comparisons between these and the Chinese taxa. DNA sequence data for all the species of Amanitaceae in China and keys for identification of the species are provided.

Keywords

Amanitaceae Combinations Novel taxa Phylogeny Species diversity 

Introduction

Amanitaceae E. J. Gilbert (Gilbert 1940) (Basidiomycota) is a cosmopolitan family comprising important species (Beeli 1931, 1935; Gilbert 1940, 1941a; Boedijn 1951; Corner and Bas 1962; Bas 1969, 1978; Reid 1978, 1980; Reid and Eicker 1991; Ridley 1991; Tulloss et al. 1995; Yang 1997, 2005, 2015; Galli 2001; Guzmán and Ramírez-Guillén 2001; Neville and Poumarat 2004, 2009; Tulloss 2005; Imazeki et al. 2011). Some edible species are economically important and are consumed in many parts of the world. For example, Amanita caesarea (Scop.) Pers. is common in European markets (Galli 2001; Pegler 2002; Román and Boa 2004), while A. caojizong Zhu L. Yang, Yang-Yang Cui and Qing Cai, reported as A. manginiana sensu W. F. Chiu (Chiu 1948) by Yang (1997, 2005, 2015), is considered a good edible mushroom in East Asia. In contrast, some species are poisonous or even deadly (Yang 2005, 2015; Deng et al. 2011; Roberts et al. 2013; Cai et al. 2014, 2016; Chen 2014; Chen et al. 2014). For example, in the period from 2000 to 2014, A. exitialis Zhu L. Yang & T.H. Li is known to have caused 45 fatalities in Guangdong, China (Yang and Li 2001; Deng et al. 2011; Chen et al. 2014, 2016).

Most species of Amanitaceae are ecologically important ectomycorrhizal taxa, and plants of more than 10 families (including Caesalpiniaceae, Casuarinaceae, Dipterocarpaceae, Fagaceae, Myrtaceae, Nothofagaceae, Pinaceae and others) are known to form symbiotic associations with Amanitaceae (Beeli et al. 1935; Reid 1980; Pegler and Shah-Smith 1997; Wood 1997; Yang 1997, 2005; Davison et al. 2017). However, some species are probably saprotrophic, e.g. Amanita pruittii A. H. Sm. ex Tulloss, J. Lindgr. & Arora, A. vittadinii (Moretti) Vittad., Catatrama costaricensis Franco-Mol., Limacella glioderma (Fr.) Maire, Limacellopsis asiatica Yang et al., Myxoderma ochraceoluteum (P.D. Orton) Yang et al. (Bas 1969; Franco-Malano 1991; Wolfe et al. 2012; Tulloss et al. 2014; Redhead et al. 2016; Yang et al. 2018).

Several genera, including Amanita Pers., Lepiota (Pers.) Gray, Limacella Earle, Pluteus Fr., Rhodotus Maire, Termitomyces R. Heim and Volvariella Speg. have been historically arranged in Amanitaceae (Singer 1962b; Moser 1967; Hawksworth et al. 1995). However, Kühner (1980), Pouzar (1983) and Singer (1986) included only Amanita and Limacella in the family.

Molecular phylogenetic analyses inferred from nucleotide sequences of the nuclear ribosomal RNA large subunit (nrLSU) suggested that the Amanitaceae comprised Amanita, Catatrama Franco-Mol. and Limacella, but the monophyly of the family was not statistically supported (Moncalvo et al. 2002; Justo et al. 2010). Vizzini et al. (2012) and Redhead et al. (2016) removed A. vittadinii (Moretti) Vittad. and its allies out of Amanita, and treated them as an independent genus, Aspidella E.-J. Gilbert or Saproamanita Redhead et al. Recently, Yang et al. (2018) suggested that Amanitaceae should include five genera, viz. Amanita, Catatrama, Limacella, Limacellopsis Zhu L. Yang et al. and Myxoderma Kühner. However, multi-locus phylogenetic studies with broader taxon sampling are needed to understand relationships between these genera.

Amanita harbors 95% of the species in the family Amanitaceae. Since the establishment of the genus by Persoon in 1797, many mycologists have contributed to the taxonomy of Amanita (Gray 1821; Roze 1876; Earle 1909; Beeli 1931, 1935; Veselý 1933; Gilbert 1940, 1941a; Konrad and Maublanc 1948; Corner and Bas 1962; Moser 1967; Jenkins 1986; Kumar et al. 1990; Yang 1997, 2005, 2015; Galli 2001; Neville and Poumarat 2004). Based on exclusively morphological studies, the genus Amanita was historically split into several genera or various subdivisions (Veselý 1933; Gilbert 1941a; Konrad and Maublanc 1948; Singer 1951; Moser 1967; Garcin 1984; Singer 1986). For example, Gray (1821) proposed a novel genus, Vaginata Gray, to accommodate the species of Amanita lacking an annulus. Roze (1876) proposed Amanitopsis Roze to harbor these exannulate Amanita species, while Earle (1909) proposed the genus Amanitella Earle to accommodate the exannulate Amanita taxa with fragile volval remnants. To date, ca. 50 sectional names have been introduced within Amanita, which has caused much controversy and confusion in the taxonomy of the genus.

Corner and Bas (1962), and Bas (1969), proposed splitting Amanita into two subgenera and six sections (sect. Amanita and sect. Vaginatae (Fr.) Quél. in subgen. Amanita; sect. Amidella (E. J. Gilbert) Konrad & Maubl., sect. Lepidella, sect. Phalloideae (Fr.) Quél. and sect. Validae (Fr.) Quél. in subgen. Lepidella Beauseigneur) by means of selection of lectotypes and treatment of later sectional names as typonymous synonyms of earlier sectional names. These nomenclatural and taxonomic treatments made significant contributions to the taxonomy of Amanita and were accepted by many mycologists (Jenkins 1977, 1986; Hongo 1982; Pegler 1983, 1986; Imazeki and Hongo 1987; Mao 1990; Ridley 1991; Tulloss et al. 1992b, 1995; Fraiture 1993; Pegler and Shah-Smith 1997), but the subgeneric classification remained controversial (Moser 1967; Garcin 1984; Singer 1986).

Based on morphological characters, phylogenetic data and basidioma development of Amanita, Yang (1997) proposed to split Amanita into two subgenera and seven sections (subgen. Amanita, including sect. Amanita, sect. Caesareae Singer ex Singer and sect. Vaginatae; and subgen. Lepidella, including sect. Amidella, sect. Lepidella, sect. Phalloideae and sect. Validae), keeping sect. Caesareae and sect. Vaginatae as two separate sections. Weiß et al. (1998), Drehmel et al. (1999), Oda et al. (1999) and Zhang et al. (2004) used nucleotide sequences of the internal transcribed spacers (ITS) or nrLSU to investigate the phylogenetic framework of Amanita, including 49, 39 and 53 species respectively, and these studies in principle supported the classification of Amanita as proposed by Yang (1997). However, only a limited number of species, and only nucleotide sequences from the two non-protein coding regions were studied.

To date, ca. 1000 species have been described worldwide in the family Amanitaceae, but only ca. 600 are treated as good species (Yang et al. 2018). In China, about 130 taxa of Amanitaceae have been reported (Yang 1997, 2005, 2015; Zhang et al. 2010; Deng et al. 2014, 2016; Li and Cai 2014; Ariyawansa et al. 2015; Li et al. 2015; Cai et al. 2016; Liu et al. 2017), with over 20 putative new taxa listed but without formal descriptions (Yang 1997, 2005, 2015). Moreover, according to our previous work (Cai et al. 2014, 2016; Ariyawansa et al. 2015), morpho-species often harbor more than one phylogenetic species. For example, Chinese collections of the lethal Amanita with gray basidiomata were commonly treated as A. fuliginea Hongo based on morphological data (Yang 2005). However, our studies indicate that the species complex includes at least four species: A. fuliginea, A. fuligineoides P. Zhang & Zhu L. Yang, A. griseorosea Qing Cai et al. and A. subfuliginea Qing Cai et al. (Zhang et al. 2010; Cai et al. 2016). Therefore, species diversity of Chinese Amanitaceae has probably been considerably underestimated.

The aims of this study are (1) to construct a phylogeny of the Amanitaceae with multi-locus data; (2) to present higher levels of classification within the family Amanitaceae; and (3) to document the Chinese species of Amanitaceae using morphological characters, molecular phylogenetic evidence and ecological data.

Materials and methods

Sample collection

Samples collected in southwestern, central, southeastern, northeastern and northwestern China over the last two decades were examined. Whenever possible, samples from other parts of the world (South and Southeast Asia, Europe, North and Central America, New Zealand, Australia and Africa) were also included. Voucher specimens were deposited in the Herbarium of Cryptogams at the Kunming Institute of Botany, Chinese Academy of Sciences (KUN-HKAS). Additional collections on loan from other herbaria were studied. Loans included those from the Herbarium Bogoriense, Research Centre for Biology, Cibinong, Indonesia (BO); the Cornell Plant Pathology Herbarium, Cornell University, New York, USA (CUP); the Canadian National Mycological Herbarium, Ottawa, Ontario, Canada (DAOM); the Field Museum of Natural History, Chicago, Illinois, USA (F); the Farlow Herbarium of Harvard University Cambridge, Massachusetts, USA (FH); the Forest Research Institute of Malaysia (FRIM), the Mycological Herbarium of the Guangdong Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou, China (GDGM); the Herbarium of Mycology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China (HMAS); the Herbarium of Mycology, Jilin Agricultural University, Changchun, Jilin, China (HMJAU); the Herbarium of Northeast China, Institute of Applied Ecology, Academia Sinica, Shenyang, Liaoning, China (IFP); the herbarium of the Royal Botanic Gardens, Kew, England, UK (K); the Kanagawa Prefectural Museum of Natural History, Odawara, Kanagawa, Japan (KPM); the Rijksherbarium, Leiden, Netherlands (L); the Herbarium Marburgense, Philipps-Universität Marburg, Marburg, Germany (MB); the National Herbarium of Victoria, Australia (MEL); the Herbarium of Mycology, Hunan Normal University, Hunan, China (MHHNU); the New Zealand Fungal Herbarium, Auckland, New Zealand (PDD); the Western Australian Herbarium, Perth, Western Australia, Australia (PERTH); the Private Herbarium Rooseveltensis Amanitarum, USA (RET); the Herbarium of the Hokkaido University Museum, Sapporo, Hokkaido, Japan (SAPA); the Mycological Herbarium of Tropical Botanic Garden and Research Institute, Trivandrum, Kerala, India (TBGT); the Forestry and Forest Products Research Institute, Kukizaki, Ibaraki, Japan (TF); the Herbarium of the National Museum of Natural Science, Taichung, Taiwan, China (TNM); the Herbarium of the National Science Museum, Tokyo, Japan (TNS); the Herbarium of the Institute of Biology and Soil Science, Russian Academy of Science, Russia (VLAM); and the Herbarium of the Institute of Botany at the University of Vienna, Austria (WU). Acronyms of the herbaria follow Thiers (2018).

DNA extraction, PCR, sequencing and nucleotide alignment

Protocols for DNA extraction, PCR amplification and sequencing follow those described in Cai et al. (2014) and references therein. The following primer pairs were used for PCR amplification and sequencing: ITS1F and TIS4 were used for the internal transcribed spacers 1 and 2 with the 5.8S rDNA (ITS) (White et al. 1990; Gardes and Bruns 1993); LROR and LR5 (Vilgalys and Hester 1990) for the D1–D3 domains of the nuc 28S rDNA (nrLSU); EF1-983F and EF1-1567R (Rehner and Buckley 2005) for the translation elongation factor 1-α (tef1-α); Am-6F and Am-7R, and Am-b-tubulin F and Am-b-tubulin R (Cai et al. 2014) for the RNA polymerase II second largest subunit (rpb2) and the beta-tubulin (β-tubulin) gene fragments, respectively. The newly generated sequences were assembled and edited using SeqMan (DNA STAR package; DNAStar Inc., Madison, WI, USA) with generic-level identities for sequences confirmed via Blast queries of GenBank.

In total, ca. 890 samples of Amanitaceae were included in the molecular phylogenetic analyses. These vouchers are not all shown in the phylogenetic trees, but the detailed information about them, including GenBank accession numbers, is provided in Table S1. Four datasets were assembled for this study. Dataset I (nrLSU/rpb1/rpb2) was used to test the monophyly of Amanitaceae. This dataset consisted of 200 taxa of ca. 51 families of Agaricales, with certain species of Serpulaceae, Hygrophoropsidaceae and Coniophoraceae selected as outgroup taxa (Matheny et al. 2006). Dataset II (nrLSU/rpb2/tef1-α/β-tubulin) was a concatenated dataset, including only one to three representative samples per species, and was used to infer the phylogeny of Amanitaceae. In this dataset, Melanoleuca verrucipes (Fr.) Singer, Melanoleuca communis Sánchez-García & Cifuentes, Pluteus hongoi Singer and Pluteus romellii (Britzelm.) Sacc. were used as outgroups (Garnica et al. 2007; Justo et al. 2011; Zhao et al. 2017). Dataset III (ITS) was employed to infer the phylogenetic relationships between the Amanita species from China and those of other areas, because GenBank contains a large amount of ITS sequence data from the genus. These related ITS sequences were retrieved from GenBank by using the genus search tool in emerencia (Nilsson et al. 2005), and sequences with very short lengths and redundant identical sequences from the same phylogenetic species were later excluded from the final analyses. Given that the ITS sequences were too variable to obtain reliable alignment across the whole genus, these sequences were divided into different datasets according to sections the samples came from. Dataset IV (nrLSU/rpb2/tef1-α) was used to delimit the species boundaries of Amanitaceae. GenBank accession numbers and related information regarding the vouchers included in the datasets were listed in Tables S1–S3.

Alignments were constructed separately for each of the gene fragments using MAFFT v7.310 (Katoh and Standley 2013), optimized using BioEdit v7.0.9 (Hall 1999), and then concatenated using SequenceMatrix v1.7.8 (Vaidya et al. 2011). Unsampled gene regions were coded as missing data. In the three concatenated datasets (Dataset I, II and IV), all introns of rpb1, rpb2, tef1-α, and β-tubulin were excluded because of the difficulty in alignment.

Phylogenetic analyses and species delimitation

The best partition schemes and evolutionary models for all the datasets were selected using PartitionFinder v2.1.1 (Lanfear et al. 2016). Prior to partition scheme selection, the data block was manually defined. The exons of rpb1, rpb2, tef1-α and β-tubulin were separated into three blocks by the first, second and third codons, respectively. ITS were divided into three blocks, ITS1, ITS2 and 5.8S, and nrLSU was treated as a single block. The best partition schemes and best fit models for the datasets are summarized in Tables S4–S7. For the concatenated datasets, single-locus phylogenies were constructed to detect incongruence among individual genes using the maximum likelihood (ML) method. As no conflicts were found (e.g. well-supported differences in the topology; Figs. S2–S3 and S15–S17), these sequences were then concatenated for further multi-locus phylogenetic anlayses. The ML analysis was performed using RAxML v7.9.1 (Stamatakis 2006), implementing the GTR + GAMMA + I or GTR + GAMMA model, and executing a rapid bootstrapping (MLB) with 1000 replicates. The final alignments have been submitted to TreeBase (No. 22967).

To infer species boundaries in Amanitaceae, two methods were employed. The first was the Genealogical Concordance Phylogenetic Species Recognition (GCPSR) criterion (Taylor et al. 2000), which has proven to be a good tool for species recognition in fungi (Giraud et al. 2008; Vialle et al. 2013; Han et al. 2018). In this study, the phylogenetic species were defined according to the two GCPSR-based criteria (Dettman et al. 2003): (1) genealogical concordance: the lineage is well supported in the majority (3/4) of the single-locus genealogies; (2) genealogical nondiscordance: the lineage is well-supported by at least one single-locus genealogy (MLB ≥ 70%), and is not contradicted in any other single-locus genealogy at the same level of support. However, for those closely related species, which are in the initial stages of divergence, it could be difficult to recognize using GCPSR method, since genes can differ in their evolutionary histories due to incomplete lineage sorting, horizontal gene transfer and population structure (Degnan and Rosenberg 2009; Stewart et al. 2014). In addition, species with only a single collection could not be analyzed using GCPSR. Therefore, a coalescent-based species delimitation method, Poisson Tree Processes (PTP) (Zhang et al. 2013), was also employed. Species delimitation based on coalescent models attempts to incorporate the process of lineage sorting and the incongruent genomic regions into phylogenetic analyses (Carstens and Knowles 2007). The PTP method estimates the mean expected number of substitutions per site between two branching events, using the branch length information of a phylogeny and then implements two independent classes of poisson processes (intra and inter specific branching event), before clustering the phylogenetic trees according to the results. Our analysis was carried out on the web server for PTP (http://species.h-its.org/) using the RAxML tree.

Morphological studies

Macroscopic characters were described from fresh basidiomata, detailed field notes and photographs. Color codes indicated in the descriptions, e.g. “2C2–4’’, follow Kornerup and Wanscher (1981). Microscopic structures were observed with light microscopy from the dried specimens after sectioning and mounting in 5% KOH solution, and where necessary, staining with Congo Red. Melzer’s reagent was used to test the amyloidy of the basidiospores. In the descriptions of basidiospores, the abbreviation (n/m/p) means n basidiospores measured from m basidiomata of p collections; sizes for basidiospores are given using a range notation of the form (a–) b–c (–d), the range b–c contains a minimum of 90% of the measured values, extreme values (a, d) are given in parentheses; Q is used to present “length/width ratio” of a spore in side view; Qm is the mean Q of all basidiospores ± sample standard deviation. Statistical analysis of the measurements for the basidiospores was conducted in SigmaPlot 10.0. The descriptive terms follow Bas (1969), Yang (1997, 2005, 2015), Yang et al. (2004), and Cai et al. (2016).

The generic names applied in this paper are abbreviated as follows: Amanita = “A.”, Aspidella = “Asp.”, Catatrama = “C.”, Limacella = “L.”, Limacellopsis = “Lim.”, Myxoderma = “M.”, Saproamanita = “S.” and Pinus = “P.

Results

Phylogenetic analyses

In Dataset I (nrLSU/rpb1/rpb2), 398 sequences from 200 species were included. The length of the dataset was 2424 bp after alignment and exclusion of 942 bp of introns, with 1021 bp for nrLSU, 1611 bp for rpb1, and 734 bp for rpb2. The highest-scoring RAxML tree is shown with bootstrap support values in Fig. S1. Our analysis strongly supported the monophyly of Amanitaceae (MLB = 94) and recognized five genera within the family: Amanita, Catatrama, Limacella, Limacellopsis and Myxoderma.

Dataset II (nrLSU/rpb2/tef1-α/β-tubulin) comprised 1004 sequences from 377 representative samples. The aligned length was 2458 bp after exclusion of 227 bp introns, with lengths of 1063, 697, 463 and 235 bp for nrLSU, rpb2, tef1-α, and β-tubulin, respectively. Our results suggested that Amanita was sister to the rest of Amanitaceae with weak support (MLB = 52%), while Catatrama and Limacella had a close relationship with strong support (MLB = 100%) (Fig. 1). Within Amanita, three subgenera and 11 sections were recovered with high bootstrap support. The subgen. Lepidella, comprising only one section, sect. Lepidella, occupied a basal position to Amanita. Amanita subgen. Amanita comprised sect. Amanita, sect. Amarrendiae, sect. Caesareae and sect. Vaginatae. Though the phylogenetic relationships between the four sections were unresolved, sect. Amanita clustered together with sect. Amarrendiae with low support (MLB = 52%). In subgen. Amanitina, sect. Roanokenses and sect. Amidella occupied the basal and sub-basal positions. In the remaining four sections, sect. Validae and sect. Strobiliformes were sister to each other with moderate support (MLB = 80%) (Fig. 1).
Fig. 1

Phylogeny of Amanitaceae inferred from the Dataset II (nrLSU/rpb2/tef1-α/β-tubulin) using the maximum likelihood (ML) analysis. Bootstrap values over 50 are shown above or beneath the individual branches. Sequences from type collections are marked with (T) and new species are highlighted in red. a An overview of the complete ML tree. b The complete ML tree

Dataset III (ITS) included 701 sequences of Amanita and was divided into nine sub-datasets according to taxonomic sections (sect. Amanita, sect. Arenariae, sect. Caesareae, sect. Phalloideae, sect. Roanokenses, sect. Vaginatae and sect. Validae); A. sect. Amarrendiae, sect. Lepidella and the other four genera were not included as there were few available ITS sequences from representative taxa in GenBank. The phylogenetic relationships between species inferred from ITS sequences data were somewhat different from those inferred from Dataset II (Fig. 1) perhaps because more species were included in these analyses (Figs. S6–S14).

Species recognition

Dataset IV (nrLSU/rpb2/tef1-α) included 1459 sequences, and had an aligned length of 2208 bp after exclusion of 227 bp of introns. According to the two GCPSR-based criteria, a total of 155 phylogenetic species were recognized, of which 132 fulfilled the criteria. The remaining 23 were phylogenetic species, represented by single samples, and were delimited since they were highly divergent from their sister groups (e.g. A. hamadae, A. hemibapha, A. neocinctipes, and A. zambiana) (Fig. 2). With the PTP method, 199 phylogenetic species were delimited (Fig. 2). More phylogenetic species were recognized with the second method than the first one, probably because the intraspecific divergences of several phylogenetic species recognized in the GCPSR method were large, and these species were therefore divided into different phylogenetic species using the PTP method. Examples of such species include A. avellaneosquamosa, A. chiui, A. farinosa, A. minutisquama, A. miculifera, A. oberwinklerana, A. ovalispora, and A. pseudoporphyria. In addition, there were another 30 phylogenetic species represented by only a single sample. These were defined together with their sister groups as single phylogenetic species according to the GCPSR criterion, as they were closely related to the phylogenetic species with more than one sample. However, the PTP method defines these as different phylogenetic species. An example is that of A. muscaria, A. ibotengutake, and A. flavopantherina, which formed a monophyletic group and were recognized as one phylogenetic species (represented by A. flavopantherina) by the GCPSR method, but were defined as three different phylogenetic species by the PTP method (Fig. 2).
Fig. 2

Species recognition of Amanitaceae inferred from the Dataset IV (nrLSU/rpb2/tef1-α) using the maximum likelihood (ML) analysis. Bootstrap values over 50 are shown above or beneath the individual branches. Sequences from type collections are marked with (T) and new species are highlighted in red

Finally, 178 phylogenetic species were accepted by considering the two methods together with morphological and ecological evidence (Fig. 2). Although the majority of the species corresponded to the phylogenetic species, a few were represented by more than one phylogenetic species. For example, A. aspericeps, A. oberwinkelerana, A. pallidorosea, and A. sepiacea were shown to be two phylogenetic species by both the GCPSR and PTP methods, but they were finally treated as a single species because it was difficult to distinguish the two closely related lineages based on both morphological studies and ecological evidence. In contrast, A. phalloides and A. subjunquillea were shown to be a single phylogenetic species in the two analyses, but they were delimited as two different species because of their differences in morphology and geographic distribution.

In the aforementioned phylogenetic species, 140 of them were from China which consisted of 87 known species, 46 novel taxa and seven putatively new species (Fig. 2). In addition, due to a lack of rpb2 and tef1-α sequences used for species delimitation, another four novel species (A. parvivirginea, A. innatifibrilla, A. lignitincta and A. pannosa) and 18 known taxa from China were excluded in the phylogenetic species recognition (Fig. 2). However, they were included in the phylogenetic analyses (Fig. 1), and were delimited by their distinct morphological characters and high divergence from the sister groups based on the nrLSU sequences. Ultimately, 162 phylogenetic species from China were recognized. Among them, 155 were documented in this study, comprising 105 known species and 50 new taxa. The remaining seven putatively new species, A. cf. angustilamellata (HKAS 89451), A. cf. incarnatifolia (HKAS 92048/HKAS 100579/HKAS 100633), A. cf. vestita (HKAS 80175), A. aff. princeps (RET 357-5), Amanita sp. (HKAS 89019/HKAS 91096/HKAS 91972/HKAS 101389/HKAS 101390), Amanita sp. (HKAS 75475), and A. cf. xanthogala (HKAS 83444/HKAS 84707), are not described here, because it is difficult to distinguish them from their related species, and more observations are be needed.

Morphological observations

In this work, 1190 specimens, consisting of 1008 Chinese specimens and 182 external collections, were studied. The Chinese specimens represent 162 taxa and ten insufficiently known taxa. The 162 Chinese taxa include 50 novel species and a total of 112 known taxa (104 species, four varieties, and four forms). Among the 162 Chinese taxa, 155 of them were supported with molecular phylogenetic evidence, morphological characters and ecological data, while seven of them were not included in phylogenetic analyses due to poor conditions of these specimens. In addition, the ten insufficiently known taxa of Amanita recorded from China were briefly listed at the end of the taxonomic part of the genus Amanita and were not formally described due to lack of available materials, and are in need of further investigation. The external collections represent 26 species, including a novel species, namely Catatrama indica. They were studied in order to make morphological and molecular phylogenetic comparisons with the Chinese taxa.

Taxonomy

Based on our multi-locus phylogenetic, morphological and ecological data, we provide details of the higher levels of classification within the family Amanitaceae and treat the species of Amanitaceae from China below.

System of Amanitaceae

Amanitaceae E. J. Gilbert 1940

Type: Amanita Pers. 1797
  1. I.

    Amanita Pers. 1797

    Lectotype: A. muscaria (L.) Lam. 1783
    1. i.

      Subgen. Amanita

      Type: A. muscaria (L.) Lam. 1783
      1. 1.

        Sect. Amanita

        Type: A. muscaria (L.) Lam. 1783

         
      2. 2.

        Sect. Amarrendiae (Bougher & Lebel) Zhu L. Yang, Yang-Yang Cui, Qing Cai & Li-Ping Tang

        Type: A. oleosa (Bougher & T. Lebel) Justo 2010

         
      3. 3.

        Sect. Caesareae Singer ex Singer 1951

        Lectotype: A. caesarea (Scop.) Pers. 1801

         
      4. 4.

        Sect. Vaginatae (Fr.) Quél. 1872

        Lectotype: A. vaginata (Bull.: Fr.) Lam. 1783

         
       
    2. ii.

      Subgen. Amanitina (E. J. Gilbert) E. J. Gilbert 1941

      Type: A. phalloides (Vaill.: Fr.) Link 1833
      1. 5.

        Sect. Amidella (J. E. Gilbert) Konrad & Maubl. 1948

        Type: A. volvata (Peck) Lloyd 1898

         
      2. 6.

        Sect. Arenariae Zhu L. Yang, Yang-Yang Cui & Qing Cai

        Type: A. arenaria (O. K. Mill. & E. Horak) Justo 2010

         
      3. 7.

        Sect. Phalloideae (Fr.) Quél. 1872

        Lectotype: A. phalloides (Fr.: Fr.) Link. 1833

         
      4. 8.

        Sect. Roanokenses Singer ex Singer 1962

        Type: A. roanokensis Coker 1927

         
      5. 9.

        Sect. Strobiliformes Singer ex Zhu L. Yang, Yang-Yang Cui & Qing Cai

        Type: A. strobiliformis (Paul. ex Vitt.) Bertillon 1866

         
      6. 10.

        Sect. Validae (Fr.) Quél. 1972

        Lectotype: A. excelsa (Fr.) Bertillon 1866

         
       
    1. iii.

      Subgen. Lepidella Beauseigneur 1926

      Type: A. vittadinii (Moretti) Vittad. 1826
      1. 11.

        Sect. Lepidella Corner & Bas 1962

        Type: A. vittadinii (Moretti) Vittad. 1826

         
       
     
  2. II.

    Catatrama Franco-Mol. 1991

    Type: C. costaricensis Franco-Mol. 1991

     
  3. III.

    Limacella Earle 1909

    Type: L. delicata (Fr.) Earle ex Konr. & Maubl. 1930

     
  4. IV.

    Limacellopsis Zhu L. Yang et al. 2018

    Type: Lim. guttata (Pers.) Zhu L. Yang et al. 2018

     
  5. V.

    Myxoderma Kühner 1926

    Type: M. illinitum (Fr.) Kühner 1926

     
Amanitaceae E. J. Gilbert, Iconogr. mycol., Suppl. I 27(fasc. I): 63 (1940)

Type: Amanita Pers., Tent. disp. meth. fung. (Lipsiae): 65 (1797).

Diagnosis: Basidioma mostly agaricoid, occasionally sequestrate; lamellar trama bilateral; subhymenium composed of inflated cells; context of stipe (if present) with terminal inflated cells (acrophysalidic); basidiospores colorless or nearly so and hyaline, smooth to echinulate, amyloid or inamyloid.

Key to the genera of Amanitaceae

  1. 1.

    1. Basidioma agaricoid……….2

     
  2. 1.

    Basidioma sequestrate, stipitate or astipitate, with schizohymenial in development………Amanita

     
  3. 2.

    Volval remnants on pileus often present, sometimes absent; pileipellis a cutis or ixo-cutis composed of non-gelatinized or gelatinized, repent filamentous hyphae……….3

     
  4. 2.

    Volval remnants on pileus absent; pileipellis an ixotrichoderm composed of erect filamentous hyphae or inflated cells……….4

     
  5. 3.

    Annulus present and obvious, or absent; stipe base inflated or not; volval remnants on stipe base usually present, occasionally absent; basidiospores often smooth, rarely asperulate………Amanita

     
  6. 3.

    Annulus present, but rudimental; stipe base bulbless; volval remnants on stipe base absent or present; basidiospores asperulate to warted………Catatrama

     
  7. 4.

    Annulus absent or not obvious; pileipellis an ixotrichoderm composed of subcylindrical terminal cells, inflated cells in short chains absent……….5

     
  8. 4.

    Annulus present and large; pileipellis an ixotrichoderm composed of inflated cells in short chains, and attenuate terminal cells………Limacellopsis

     
  9. 5.

    Stipe dry and covered with squamules; annulus non-gelatinized and rudimental………Limacella

     
  10. 5.

    Stipe glutinous to strongly viscid, glabrous or covered with squamules; annulus absent, or strongly gelatinized if present………Myxoderma

     

Amanita Pers., Tent. disp. meth. fung. (Lipsiae): 65 (1797).

Lectotype: Amanita muscaria (L.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 111 (1783).

Diagnosis: Basidioma mostly agaricoid, occasionally sequestrate, schizohymenial in development; lamellar trama bilateral; basidiospores colorless and hyaline; trama with inflated terminal cells. When the basidioma is agaricoid: lamellae free; volval remnants present; annulus present or absent; basidiospores ballistosporic, smooth, rarely asperulate; pileipellis a cutis or ixo-cutis composed of more or less gelatinized, repent filamentous hyphae. When the basidioma is sequestrate: volval remnants or peridium present; gleba loculate; basidiospores statismosporic, smooth; basidioma mainly distributed in regions with dry and warm Mediterranean climates (southern Europe, northern Africa, and Australia).

Key to the subgenera of the genus Amanita

  1. 1.

    Basidiospores inamyloid; when the basidioma is agaricoid, pileal margin striate and lamellulae truncate; when the basidioma is sequestrate, basidia thin-walled and clamps present………subgen. Amanita

     
  2. 2.

    Basidioma agaricoid……….3

     
  3. 2.

    Basidioma sequestrate………subgen. Amanitina

     
  4. 3.

    Stipe base elongated, bulbless; volval remnants scattered over the surface of stipe; basidioma saprotrophic………subgen. Lepidella

     
  5. 3.

    Stipe base with bulb or bulbless; volval remnants usually attached to bulb or basal part of stipe; basidioma ectomycorrhizal………subgen. Amanitina

     

Amanita subgen. Amanita

Type: Amanita muscaria (L.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 111 (1783).

Diagnosis: Basidioma mostly agaricoid, occasionally sequestrate; basidiospores inamyloid. When basidioma agaricoid, pileal margin striate; lamellulae truncate; clamps present or absent. When basidioma sequestrate, clamps present.

Commentary: Our multi-locus phylogenetic and morphological studies suggested that there are four sections in Amanita subgen. Amanita.

Key to the sections of Amanita subgen. Amanita

  1. 1.

    Basidioma agaricoid……….2

     
  2. 1.

    Basidioma sequestrate……….5

     
  3. 2.

    Volval remnants often presented as pyramids, cones, patches, or floccules to powder on pileal surface and base of stipe……….3

     
  4. 2.

    Volval remnants forming saccate structure on base of stipe, usually absent on pileal surface……….4

     
  5. 3.

    Stipe base bulbless or only slightly inflated; basidiospores mostly thin-walled and smooth, occasionally thick-walled and ornamented; clamps present; basidioma distributed in southern hemisphere and with Nothofagus, Eucalyptus, etc. ……………………………………sect. Amarrendiae

     
  6. 3.

    Stipe base usually with bulb, sometimes ventricose; basidiospores thin-walled and smooth; clamps present or absent; basidioma distributed worldwide and with Fagaceae, Pinaceae, etc. ………sect. Amanita

     
  7. 4.

    Annulus mostly absent, occasionally present; clamps absent………sect. Vaginatae

     
  8. 4.

    Annulus always present; clamps present………sect. Caesareae

     
  9. 5.

    Basidioma stipitate or astipitate, distributed in Australia and with Nothofagus, Eucalyptus, etc. ………sect. Amarrendiae

     
  10. 5.

    Basidioma stipitate, distributed in southern Europe and northern Africa, and with Pinus, Quercus, etc. ………sect. Caesareae

     

Amanita sect. Amanita

Type: Amanita muscaria (L.: Fr.) Lam., Encycl. Méth. Bot. (Paris) 1 (1): 111 (1783).

Diagnosis: Basidioma agaricoid; pileus with persistent volval remnants, pileal margin striate; lamellulae truncate; basal bulb present; basidiospores inamyloid.

Commentary: Most species in Amanita sect. Amanita contain neuropsychiatric toxins, including muscarine and isoxazole derivatives, which can produce clinical symptoms including nausea, emesis, stomach ache and deliration (Wieland 1973; Yang 2005; Chen et al. 2016). To date, 25 taxa of A. sect. Amanita from China are recognized in our phylogenetic analyses, including three novel species and 22 previously known taxa (18 species, two varieties and two forms). Amanita gleocystidiosa and A. pyriformis are treated as synonyms of A. sychnopyramis f. subannulata and A. orientigemmata, respectively.

Key to the species of Amanita sect. Amanita from China

  1. 1.

    Annulus absent……….2

     
  2. 1.

    Annulus present……….6

     
  3. 2.

    Pileal surface grayish, grayish brown, brownish to dark brown; volval remnants on pileal surface dirty white, gray to brown-gray……….3

     
  4. 2.

    Pileal surface yellowish, yellow to orange-yellow; volval remnants on pileal surface cream to yellow………4

     
  5. 3.

    Volval remnants on pileus mostly pyramidal, rarely conical, dirty white, grayish, gray-brown to brown; basidiospores globose to subglobose, 6.5–8.5 × 6.0–8.0 μm………A. sychnopyramis f. sychnopyramis

     
  6. 3.

    Volval remnants on pileus pulverulent to minute verrucose, gray to brown-gray; basidiospores subglobose to broadly ellipsoid, 6.5–8.0 × 5.5–7.0 μm………A. farinosa

     
  7. 4.

    Basidiospores globose to subglobose, 7.5–9.5 × 7.0–9.0 μm; basidioma dominant in tropical to subtropical broad-leaved forests……….5

     
  8. 4.

    Basidiospores broadly ellipsoid to ellipsoid; basidioma dominant in subtropical to temperate pine or mixed forests………A. melleiceps

     
  9. 5.

    Pileus 4–8 cm in diam.; basidiospores smaller, 6.0–8.0 × 6.0–7.5 μm………A. mira

     
  10. 5.

    Pileus 3–5 cm in diam.; basidiospores larger, 7.5–9.5 × 7.0–9.0 μm………A. elata

     
  11. 6.

    Basidiospores globose to subglobose, occasionally broadly ellipsoid, Q = 1.0–1.25……….7

     
  12. 6.

    Basidiospores broadly ellipsoid, ellipsoid to elongate, Q = 1.15–1.9……….13

     
  13. 7.

    Pileal surface red to orange……….8

     
  14. 7.

    Pileal surface white, dirty white, gray-brown, brown, yellow-brown to yellow……….9

     
  15. 8.

    Volval remnants on pileus yellow to orange-red; basidiospores 8.5–10.5 × 8.0–10.0 μm; clamps present………A. subfrostiana

     
  16. 8.

    Volval remnants on pileus red to orange-red; basidiospores 7.5–9.0 × 7.0–8.5 μm; clamps absent………A. rubrovolvata

     
  17. 9.

    Pileal surface yellow, yellow-brown, gray-brown, brown to dark brown; volval remnants pyramidal, verrucose to patchy, reddish brown, brown, grayish to dirty white; stipe glabrous, or covered with gray to dark gray squamules……….10

     
  18. 9.

    Pileal surface white to dirty white; volval remnants pyramidal to conical, white to dirty white; stipe surface covered with floccose to pulverulent squamules………A. concentrica

     
  19. 10.

    Pileal surface yellow, gray-brown, brown to dark brown; volval remnants patchy, verrucose to pyramidal; stipe glabrous or covered with gray to dark gray squamules……….11

     
  20. 10.

    Pileal surface yellow-brown; volval remnants pulverulent, reddish brown to brown; stipe surface covered with reddish brown to brown, pulverulent squamules………A. rufoferruginea

     
  21. 11.

    Pileal surface brownish, gray-brown, brown to dark brown; volval remnants pyramidal to conical, dirty white to grayish; annulus median; basidioma distributed in subtropical forests dominated by Fagaceae……….12

     
  22. 11.

    Pileal surface cream, yellowish to yellow; volval remnants felted to floccose, yellowish, yellow to dirty yellow; annulus subapical; basidioma distributed in subalpine forests………A. altipes

     
  23. 12.

    Volval remnants on stipe base often collar-like or shortly limbate; basidiospores relatively larger, 8.0–10.0 × 7.0–8.5 μm………A. pseudosychnopyramis

     
  24. 12.

    Volval remnants on stipe base verrucose, conical to pulverulent; basidiospores relatively smaller, 6.5–8.5 × 6.0–8.0 μm………A. sychnopyramis f. subannulata

     
  25. 13.

    Clamps present……….14

     
  26. 13.

    Clamps absent……….21

     
  27. 14.

    Basal bulb subglobose to fusiform, with clear limit between stipe and basal bulb; volval remnants on stipe base recurved arranged, or collar-like; pileal surface yellow, brown or red; pileipellis gelatinized; volval remnants on pileus white, dirty white, yellowish to grayish, composed of very abundant subglobose to broadly ellipsoid, inflated cells……….15

     
  28. 14.

    Basal bulb ellipsoid to subclavate, lacking clear limit between stipe and basal bulb; volval remnants on stipe base floccose to pulverulent; pileal surface dirty white to pale grayish; pileipellis almost non-gelatinized; volval remnants on pileus gray to gray-brown, dominantly composed of very abundant, ellipsoid to fusiform, inflated cells……….20

     
  29. 15.

    Pileal surface brownish, brown to dark brown; volval remnants on stipe base often collar-like or shortly limbate……….16

     
  30. 15.

    Pileal surface yellow, red to orange; volval remnants on stipe base often as warts arranged irregularly or in concentrate incomplete belts……….19

     
  31. 16.

    Basidioma relatively larger; pileal surface relatively darker, brown to dark brown; basidioma distributed in subalpine forests……….17

     
  32. 16.

    Basidioma relatively smaller; pileal surface relatively lighter, brownish to brown; basidioma distributed subtropical or temperate forests……….18

     
  33. 17.

    Volval remnants on pileus yellow; context yellow; annulus subapical to median, yellow with a brown edge; basidiospores broadly ellipsoid to ellipsoid, 10.0–12.0 × 8.0–10.0 μm………A. flavopantherina

     
  34. 17.

    Volval remnants on pileus white to dirty white; context white; annulus apical to subapical, white to pale brownish, lacking a distinctly colored edge; basidiospores mostly broadly ellipsoid, 9.5–12.0 × 8.0–10.0 μm………A. griseopantherina

     
  35. 18.

    Basidiospores relatively smaller, 8.0–10.0 × 6.5–7.5 μm; clamps common in all parts of basidioma; basidioma distributed in temperate regions………A. ibotengutake

     
  36. 18.

    Basidiospores relatively larger, 8.5–12.0 × 7.0–9.5 μm; clamps common only in subhymenium; basidioma distributed in subtropical regions………A. subglobosa

     
  37. 19.

    Pileal surface yellowish to yellow; volval remnants on pileus felted to patchy, sometimes pyramidal; annulus fragile and fugacious; basidiospores relatively smaller, 8.0–10.0 × 6.0–7.5 μm………A. orientigemmata

     
  38. 19.

    Pileal surface red to orange, occasionally yellow; volval remnants on pileus pyramidal, verrucose to conical, sometimes felted; basidiospores relatively larger, 9.0–12.5 × 7.0–8.5 μm………A. muscaria

     
  39. 20.

    Basidiospores 9.5–12.5 × 7.0–8.5 μm, Q = 1.25–1.53, Qm = 1.38 ± 0.09, broadly ellipsoid to ellipsoid………A. sinensis var. sinensis

     
  40. 20.

    Basidiospores 8.5–10.5 × 7.5–9.0 μm, Q = 1.06–1.21, Qm = 1.13 ± 0.05, subglobose to broadly ellipsoid………A. sinensis var. subglobispora

     
  41. 21.

    Pileal surface yellow, grayish brown, brownish to brown; volval remnants on pileus verrucose, conical to pyramidal, or felted to patchy, white, cream to gray; stipe surface covered with white to dirty white, fibrous squamules……….22

     
  42. 21.

    Pileal surface yellowish brown to orange-brown, with olivaceous tinge; volval remnants on pileus pulverulent to minute verrucose, yellowish brown to orange-brown, often with olivaceous tinge; stipe surface covered with yellowish brown to orange-brown, pulverulent squamules………A. siamensis

     
  43. 22.

    Pileal surface honey yellow to yellow at center, but yellowish at margin, lacking distinct brown tinge………A. melleialba

     
  44. 22.

    Pileal surface grayish brown to brown, with distinct brown tinge……….23

     
  45. 23.

    Basidiospores relatively narrower, 8.5–11.5 × 6.5–8.0 μm……….24

     
  46. 23.

    Basidiospores relatively broader, 8.5–11.5 × 8.0–10.0 μm………A. pseudopantherina

     
  47. 24.

    Volval remnants on stipe base conical, granular, often forming a short limbate collar between stipe and basal bulb; volval remnants on pileus with fairly abundant to abundant inflated cells………A. subparvipantherina

     
  48. 24.

    Volval remnants on stipe base floccose to granular; volval remnants on pileus with more abundant to nearly dominant inflated cells………A. parvipantherina

     

1. Amanita altipes Zhu L. Yang, M. Weiß & Oberw., Mycologia 96 (3): 636 (2004).

Pileus 4–9 cm diam., yellowish (4A2–4) to yellow (4A5–8); volval remnants on pileus floccose to felted, yellowish (4A2–4) to yellow (4A5–8); margin striate (0.1–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae truncate. Stipe 9–16 cm long × 0.5–2 cm diam., yellowish (4A2–4) to white (1A1), covered with minute, yellowish (4A2–4) to yellow (4A5–8) squamules above annulus and decorated with minute, yellowish (4A2–4) to white (1A1) squamules under annulus; context white (1A1), unchanging; basal bulb subglobose, 0.8–3.2 cm diam., with its upper part covered with patchy to verrucose, yellowish (4A2–4) to yellow (4A5–8) volval remnants. Annulus superior to subapical, cream (1A2) to yellowish (4A2–4) at upper surface, yellowish (4A2–4) at lower surface; annular edge yellow.

Basidia 40–60 × 10–16 μm, clavate, 4-spored, rarely 2-spored. Basidiospores [365/16/9] (7.5–) 8.0–10.0 (–11.5) × (7.0–) 7.5–9.5 (–10.0) μm, Q = 1.0–1.14 (–1.2), Qm = 1.07 ± 0.04, globose to subglobose, rarely broadly ellipsoid, inamyloid. Volval remnants on pileus composed of radially to irregularly arranged elements: filamentous hyphae abundant to very abundant; inflated cells abundant to very abundant. Clamps absent in all parts of basidioma.

Habit: Solitary to scattered in subalpine forests dominated by Abies and Picea; basidioma occurring in summer and autumn.

Distribution: Known from central, northwestern and southwestern China (Yang et al. 2004; Yang 2005, 2015, this study).

Specimens examined: CHINA. GANSU PROVINCE: Zhouqu County, in a subalpine forest with Abies and Betula, altitude ca. 2200 m, 16 August 2012, Xue-Tai Zhu 749 (HKAS 76598). HUBEI PROVINCE: Yichang, Shennongjia, in a subalpine forest of Picea, altitude 2500 m, 17 July 2012, Qing Cai 818 (HKAS 75570). SICHUAN PROVINCE: Jiulong County, in a subalpine forest with Abies and Picea, altitude 3802 m, 20 July 2015, Jian-Wei Liu 317 (HKAS 91125); Tibetan Autonomous County of Muli, in a subalpine forest of Abies, altitude 3700 m, 2 August 2012, Ting Guo 562 (HKAS 76254). YUNNAN PROVINCE: Deqin County, in a forest of Abies, altitude 3900 m, 12 August 2015, Jian-Wei Liu 360 (HKAS 91168); Yulong Naxi Autonomous County, Laojunshan, in a subalpine forest of Abies, altitude 3800 m, 14 August 2000, Zhu L. Yang 2915 (HOLOTYPE, HKAS 36609); same location, altitude ca. 3800 m, 1 August 2005, Zhu L. Yang 4487 (HKAS 48653); same location, in a subalpine forest of Abies, altitude ca. 3500 m, 4 September 2009, Qing Cai 225 (HKAS 58890); Shangri-La, in a subalpine forest of Abies, altitude 3750 m, 25 August 2009, Qing Cai 140 (HKAS 58805).

Commentary: Amanita altipes was described from China by Yang et al. (2004). It is well-characterized by its long stipe and dominates in forests at an altitude of 2500–4000 m. No species closely related to A. altipes were detected in A. sect. Amanita based on our multi-locus phylogenetic analysis (Fig. 1b). For detailed descriptions, comparisons with similar species, line drawings and images of A. altipes see Yang et al. (2004) and Yang (2005, 2015).

2. Amanita concentrica T. Oda, C. Tanaka & Tsuda, Mycoscience 43 (1): 81 (2002).

Pileus 7–13 cm in diam., white (1A1) to dirty white (2B1); volval remnants on pileus pyramidal to conical, ca. 1–3 mm in height and width, white (1A1) to dirty white (2B1); margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2–3), lamellulae truncate. Stipe 7–17 cm long × 0.8–1.7 cm diam., white (1A1), covered with floccose to pulverulent, white squamules; context white (1A1), unchanging; basal bulb ovoid to subglobose, 2–3 cm diam., with its upper part covered with pyramidal to verrucose, white (1A1), concentrically arranged volval remnants. Annulus subapical to submedian, persistent or fugacious.

Basidia 40–55 × 9–11 μm, clavate, 4-spored. Basidiospores [40/3/3] 7.5–9.5 (–12.5) × (6.5–) 7.0–8.0 (–10.0) μm, Q = (1.0–) 1.07–1.25 (–1.36), Qm = 1.15 ± 0.07, subglobose to broadly ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae scarce; inflated cells very abundant to nearly dominant. Volval remnants on stipe base similar to structure of volval remnants on pileus, but with irregularly arranged elements and more filamentous hyphae. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in broad-leaved or mixed forests dominated by Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2005, 2015; this study), Japan (Oda et al. 2002b; Imazeki et al. 2011), India (Bhatt et al. 2003; Semwal et al. 2006b, 2007) and Thailand (Thongbai et al. 2016).

Specimens examined: CHINA. YUNNAN PROVINCE: Jinghong, Dadugang, in a subtropical forest dominated by Fagaceae, altitude 1370 m, 5 July 2003, Fu-Qiang Yu 1093 (HKAS 44733); Mengla County, in a mixed forest with Fagaceae and Pinaceae, altitude 700 m, 9 July 2014, Xiao-Bin Liu 414 (HKAS 87061); same location, in a subtropical forest with Fagaceae, altitude ca. 700 m, 6 July 2014, Li-Hong Han 379 (HKAS 84675).

Specimens of A. gemmata examined: NETHERLANDS. SOUTH HOLLAND PROVINCE: Schoorl, West of village, in pine-plantation on non-calcareous dunes, 26 November 1994, C. Bas 8949 (L). FRIESLAND PROVINCE: Vlieland, in planted pine forests on coastal dunes, 23 October 1994, C. Bas 8942 (L).

Commentary: Amanita concentrica was described from Japan by Oda et al. (2002b). It was then reported from China, India and Thailand (Yang 2005, 2015; Semwal et al. 2007; Thongbai et al. 2016).

In our multi-locus data (Fig. 1b), A. concentrica clustered, with low support, with the A. muscaria group, which consists of A. gemmata (Fr.) Bertillon, A. muscaria (L.) Lam., A. regalis (Fr.) Michael, A. persicina (Dav.T. Jenkins) Tulloss & Geml., A. ibotengutake T. Oda, C. Tanaka & Tsuda and A. flavopantherina Yang-Yang Cui et al. However, the two specimens of A. gemmata from the Netherlands listed above differ from A. concentrica by their grayish yellow to yellowish pilei, volval remnants on the pileus dominantly composed of subradically arranged elements with very abundant to nearly dominant filamentous hyphae, volval remnants on the stipe base limbate, basidiospores [120/6/2] (9–) 9.5–12.0 (–13.0) × (6.0–) 6.5–8.5 (–9.0) μm, Q = (1.17–) 1.25–1.62 (–1.77), Qm = 1.42 ± 0.13, broadly ellipsoid to ellipsoid, and the absence of clamps.

The other species in this group differ morphologically from A. concentrica in the following ways. Amanita muscaria has a red pileus and broadly ellipsoid to ellipsoid basidiospores (9.0–12.5 × 7.0–8.5 μm) (Gilbert 1940, 1941a; Yang and Oberwinkler 1999; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015). Amanita regalis has an ocher-yellow, ocher-brown to brown pileus covered with floccose to pulverulent, yellowish to yellowish-gray volval remnants and relatively longer basidiospores (9.0–11.0 × 7.0–8.0 μm) (Gilbert 1940, 1941a; Contu 2000a; Neville and Poumarat 2004). Amanita persicina has a pastel red to light orange or melon-colored pileus covered with yellowish to tannish, floccose-fibrillose patches, cream lamellae with a pinkish tint, the context of the stipe turns yellowish when cut or bruised and the basidiospores are relatively longer (9.4–12.7 × 7.0–8.5 μm) (Jenkins 1977, 1986; Tulloss et al. 2015). Amanita ibotengutake has a yellow-brown pileus, short collar-like volval remnants on the stipe base and broadly ellipsoid basidiospores (8.0–10.0 × 6.0–7.5 μm) (Oda et al. 2002c; Yang 2015). Amanita flavopantherina has a brown pileus covered with yellow, verrucose volval remnants, collar-like volval remnants on the stipe base, broadly ellipsoid to ellipsoid basidiospores (10.0–12.0 × 8.0–10.0 μm) and is dominant in subalpine forests with Picea and Abies (see below).

For detailed descriptions, comparisons with similar species, line drawings and images of A. concentrica see Yang (2005, 2015).

3. Amanita elata (Massee) Corner & Bas, Persoonia 2 (3): 286 (1962).

Basionym: Collybia elata Massee, Bull. Misc. Inf., Kew: 73 (1914).

Basidioma small. Pileus 3–5 cm in diam., yellowish brown (4B3–5) at center, becoming yellowish (2A2–3) to white (1A1) towards margin; volval remnants on pileus floccose to pulverulent; margin striate (0.4–0.6 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 5–7.5 cm long × 0.3–0.7 cm diam., cream (1A2) to white (1A1); context white (1A1), unchanging; basal bulb top-shaped to subglobose, 1–1.7 cm diam., with its upper part covered with pulverulent to verrucose, white (1A1) to dirty white (2B1) volval remnants. Annulus often absent, occasionally present.

Basidia 28–54 × 10–14 μm, clavate, 4-spored. Basidiospores [40/2/2] 7.5–9.5 (–10.0) × 7.0–9.0 μm, Q = (1.0–) 1.03–1.19 (–1.28), Qm = 1.11 ± 0.05, globose to subglobose, inamyloid. Volval remnants on pileus composed of vertically arranged filamentous hyphae and inflated cells. Volval remnants on stipe base composed of irregularly arranged filamentous hyphae and inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical, broad-leaved forests dominated by Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from eastern and southwestern China (Yang 2015; this study), India (Adhikari 1999; Bhatt et al. 2003; Pradeep and Vrinda 2010), Malaysia (Lee 2017), Singapore (Corner and Bas 1962) and Sri Lanka (Pegler 1986).

Specimens examined: CHINA. TAIWAN PROVINCE: in a tropical forest, altitude and date unknown, Wen-Neng Zhou 6639 (HKAS 57527). YUNNAN PROVINCE: Mengla County, in a subtropical broad-leaved forest with Fagaceae and Leguminoseae, altitude 1040 m, 6 July 2014, Xiao-Bin Liu 384 (HKAS 83449).

Commentary: Amanita elata was described from Singapore by Corner and Bas (1962), and subsequently reported from China, India, Malaysia and Sri Lanka (Pegler 1986; Adhikari 1999; Bhatt et al. 2003; Pradeep and Vrinda 2010; Yang 2015; Lee 2017). It is characterized by its small basidioma, absence of clamps and an association with broad-leaved trees in tropical to subtropical forests.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. elata is closely related to A. sychnopyramis f. sychnopyramis Corner & Bas and A. sychnopyramis f. subannulata. Comparisons between A. elata and A. sychnopyramis f. sychnopyramis are given in the commentary on A. sychnopyramis f. sychnopyramis, and the differences between A. elata and A. sychnopyramis f. subannulata are given in the commentary on A. sychnopyramis f. subannulata.

For detailed descriptions and images of A. elata see Yang (2015).

4. Amanita farinosa Schwein., Schr. naturf. Ges. Leipzig 1: 79 (1822).

Basidioma small. Pileus 3–5 cm in diam., gray (1C1–2), grayish (1B1) to pale brownish (2B2–3), densely covered with pulverulent, concolorous, easily removable volval remnants; margin striate (0.4–0.6 R), non-appendiculate or sometimes slightly appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 5–8 cm long × 0.3–0.6 cm diam., white (1A1); context white (1A1), unchanging; basal bulb ovoid to subglobose, ca. 1 cm diam., with its upper part covered with pulverulent, gray (1C1–2) volval remnants. Annulus absent.

Basidia 30–42 × 9–11 μm, clavate, 4-spored. Basidiospores [300/20/14] 6.5–8.0 × 5.5–7.0 μm, Q = 1.1–1.3, Qm = 1.18 ± 0.06, subglobose to broadly ellipsoid, inamyloid. Volval remnants on pileus composed of more or less vertically to irregularly arranged elements: filamentous hyphae fairly abundant; inflated cells very abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests dominated by Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from China (Yang 2005, 2015; this study), North America (Murrill 1913; Coker 1917; Stewart and Grund 1974; Jenkins 1977, 1986; Thiers 1982; Tulloss et al. 1995), Japan (Imai 1933, 1938; Imazeki and Hongo 1987; Imazeki et al. 2011), Republic of Korea (Kim et al. 2013b), and Russian Far East (Malysheva et al. 2014).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a forest dominated by Fagaceae, altitude 939 m, 19 July 2017, Yan-Jia Hao 1429 (HKAS 100578). FUJIAN PROVINCE: Zhangping, in a broad-leaved forest, altitude 350 m, 24 August 2013, Nian-Kai Zeng 1463 (HKAS 100490). JIANGXI PROVINCE: Fuzhou, in a forest dominated by Fagaceae, altitude 40 m, 19 June 2012, Gang Wu 916 (HKAS 77088). YUNNAN PROVINCE: Changning County, in a forest of Pinus yunnanensis, altitude 1784 m, 24 July 2009, Li-Ping Tang 923 (HKAS 56880); Jiangcheng County, in a mixed forest with Fagaceae and Pinaceae, altitude 1400 m, 29 July 2008, Li-Ping Tang 507 (HKAS 54738); Jinghong, Dadugang, in a subtropical forest with Fagaceae, altitude 1300 m, 31 July 2008, Li-Ping Tang 539 (HKAS 54770); Kunming, Heilongtan, in a forest of Pinus yunnanensis, altitude 1980 m, 18 August 2008, Zhu L. Yang 5162 (HKAS 54459); Longyang, in a forest of Pinus yunnanensis, altitude 1800 m, 20 August 2009, Qing Cai 95 (HKAS 58762); same location, in a mixed forest with Fagaceae and Pinaceae, altitude ca. 1800 m, 23 July 2009, Li-Ping Tang 903 (HKAS 56860); Jingdong County, Ailaoshan, in a forest with Pinus yunnanensis and P. kesiya var. langbianensis, altitude 2350 m, 5 August 2014, Qing Cai 1234 (HKAS 83703); same location, in a forest with Fagaceae, altitude ca. 2350 m, 6 August 2014, Qing Cai 1238 (HKAS 83707); Tengchong County, in a mixed forest with Fagaceae and Pinaceae, altitude 1905 m, 20 July 2009, Li-Ping Tang 859 (HKAS 56816); same location, in a forest of Pinus armandii and Keteleeria fortunei, altitude 2010 m, 14 August 2010, Qing Cai 394 (HKAS 67958); Yongping County, in a mixed forest with Fagaceae and Pinaceae, altitude 2087 m, 31 July 2009, Li-Ping Tang 1036 (HKAS 56993). USA. NEW JERSEY, Mercer County, Hopewell township, altitude ca. 60 m, 28 August 1982, Rodham E. Tulloss 7-28-82-E (RET 215-9); Morris County, Mendham township, altitude 214 m, 27 July 1986, Mary A. King s.n. [Tulloss 7-27-86-K] (RET 079-4). MISSISSIPPI, Jackson County, Pascagoula River State Wildlife Management Area, 16 July 1987 D. C. & Rodham E. Tulloss 7-16-87-F (RET 005-7). WEST VIRGINIA, Tucker County, 3 August 1992, Rodham E. Tulloss 8-3-92-D (RET 120-5).

Commentary: Amanita farinosa was described from North America, and subsequently reported from China, Japan, the Republic of Korea and the Russian Far East (Murrill 1913; Coker 1917; Imai 1933, 1938; Stewart and Grund 1974; Jenkins 1977, 1986; Imazeki and Hongo 1987; Thiers 1982; Tulloss et al. 1995; Yang 2005, 2015; Imazeki et al. 2011; Kim et al. 2013b; Malysheva et al. 2014). The descriptions above were based on the Chinese materials cited here. We also studied four collections from the USA as listed in specimens examined, and the basidiospores of these specimens were [80/4/4] 6.5–8.0 (–10) × (5.0–) 5.5–7.0 (–8.0) μm, Q = (1.05–) 1.12–1.34 (–1.4), Qm = 1.22 ± 0.08, often subglobose to broadly ellipsoid, or rarely ellipsoid.

Our phylogenetic data (Figs. 1b, S6) indicate that collections of A. farinosa from China and the USA form a monophyletic clade within A. sect. Amanita. Since no significant morphological differences were found between the Chinese collections and those from the USA, we treat the Chinese Amanita as A. farinosa. For detailed descriptions, comparisons with similar species and images of A. farinosa see Yang (1997, 2005, 2015).

5. Amanita flavopantherina Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 3a, b, 4.
Fig. 3

Fresh basidiomata of novel species in Amanita sect. Amanita and A. sect. Caesareae. a–b A. flavopantherina (TYPE, HKAS 82613); c–d A. griseopantherina (TYPE, HKAS 83560); e A. pseudopantherina (HKAS 83636); f–g A. alboumbelliformis (TYPE, HKAS 83448); h A. fuscoflava (TYPE, HKAS 59800); i A. ochracea (HKAS 48548); j A. pseudoprinceps (TYPE, HKAS 97523); k–l A. rubroflava (TYPE, HKAS 83089); m–n A. squarrosipes (TYPE, HKAS 76359); o A. subhemibapha (TYPE, HKAS 96847). Bars: 2 cm

Fig. 4

Microscopic features of Amanita flavopantherina (TYPE, HKAS 82613). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 824957

Etymology: flavopantherina, from flavus = yellow, and pantherina = Amanita pantherina, is proposed because this species is similar to European A. pantherina, but has yellow squamules on pileus.

Type: CHINA. YUNNAN PROVINCE: Shangri-la, in a subalpine forest dominated by Picea and Quercus, altitude 3630 m, 3 September 2013, Bang Feng 1507 (TYPE, HKAS 82613, GenBank Acc. No.: ITS = MH508355, nrLSU = MH486519, tef1-α = MH508795, rpb2 = MH485989, β-tubulin = MH485512).

Basidioma (Fig. 3a, b) medium-sized to large. Pileus 5–13 cm diam., plano-convex to applanate, lacking an obviously depression or umbo at center, yellow-brown (4B3–6), brownish (3C3–5), brown (4C3–6) to dark brown (4D4–8), darker at center, but becoming paler towards the margin; volval remnants on pileus pyramidal, subverrucose to subconical, ca. 2–5 (–10) mm diam., yellowish (2A3–5), densely arranged over disk; margin slightly striate (ca. 0.05–0.1 R), non-appendiculate; trama white (1A1) to yellowish (2A3–5), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe 8–23 cm long × 1–3.5 cm diam., cylindrical and slightly tapering upwards, with apex slightly expanded, white (1A1), pale yellowish (1A2–4) to yellowish (2A3–5), glabrous or covered with concolorous, floccose squamules; context white (1A1) to yellowish (2A3–5); basal bulb subglobose to fusiform, 1.5–3.5 cm diam., white (1A1), pale yellowish (1A2–4) to yellowish (2A3–5); volval remnants on stipe base floccose, arranged in belts on the lower part of stipe, and often formed a collar-like or shortly limbate volva on limit between stipe and basal bulb, white (1A1), pale yellowish (1A2–4) to yellowish (2A3–5). Annulus persistent, subapical to median, pendant from attachment ca. 1.5–4 cm below apex of stipe, white (1A1), pale yellowish (1A2–4) to yellowish (2A3–5), membranous, with a yellow (3A5–7) to brown (4C6–8) edge. Odor unpleasant.

Lamellar trama bilateral. Mediostratum 35–50 μm wide, composed of abundant subglobose, ellipsoid to clavate inflated cells (15–90 × 15–50 μm); filamentous hyphae fairly abundant to abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid inflated cells (35–50 × 15–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant, 4–8 μm wide. Subhymenium (Fig. 4a) 30–70 μm thick, with 2–3 layers of ovoid, subglobose, fusiform, ellipsoid or irregular cells, 10–30 × 8–17 μm. Basidia (Fig. 4a) 50–70 × 12–16 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 4b) [120/6/3] 10.0–12.0 (–13.0) × (7.0–) 8.0–10.0 (–11.0) μm, Q = (1.09–) 1.20–1.47 (–1.64), Qm = 1.34 ± 0.10, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (15–30 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–10 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 80–175 μm thick; upper layer (30–75 μm thick) gelatinized, composed of radially, thin-walled, colorless, filamentous hyphae 2–8 μm wide; lower layer (40–100 μm thick) composed of radially and compactly arranged, filamentous hyphae 3–8 μm wide, colorless to brown; vascular hyphae scarce. Volval remnants on pileus (Fig. 4c) composed of somewhat vertically to irregularly arranged elements: filamentous hyphae scarce to scattered, 3–7 μm wide, subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to dominant, globose, subglobose, fusiform to ellipsoid, sometimes irregular, 40–80 × 20–55 μm, nearly colorless, slightly thick-walled (≤ 1 μm), terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base dominantly composed of two parts intergrading to each other. Outer part composed of irregularly arranged elements: filamentous hyphae abundant to very abundant, 3–7 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, becoming fairly abundant towards inner part, fusiform to ellipsoid, sometimes elongate, 30–110 × 20–60 μm, colorless, terminal or in chains of 2–3; vascular hyphae scarce. Inner part dominantly composed of irregularly arranged elements: filamentous hyphae very abundant to nearly dominant, 3–10 (–15) μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce, scattered to fairly abundant, ellipsoid to elongate, 60–110 × 30–50 μm, colorless, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of outer part; inner surface similar to the structure of inner part but gelatinized and with more filamentous hyphae. Stipe trama composed of longitudinally arranged, clavate terminal cells, 90–380 × 15–50 μm; filamentous hyphae scattered to abundant, 3–13 μm wide; vascular hyphae scarce. Annulus dominantly composed of radially arranged elements: filamentous hyphae very abundant, 2–14 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant, subglobose, fusiform, ellipsoid to elongate 25–120 × 10–50 μm, colorless, terminal or in chains of 2–3; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in subalpine forests dominated by Abies and Picea; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Shangri-la, in a subalpine forest dominated by Picea and Rhododendron, altitude 3650 m, 24 August 2009, Qing Cai 128 (HKAS 58795). SICHUAN PROVINCE: Daocheng County, in a subalpine forest with Abies, Picea and Spiraea, altitude 4055 m, 27 August 2006, Zai-Wei Ge 1460 (HKAS 51045).

Commentary: Amanita flavopantherina is easily distinguished by its brown to dark brown pileus covered with pyramidal to subverrucose, yellowish volval remnants, and with short striations on the margin, a white to yellow context, a white to yellow stipe with a collar-like, shortly limbate volva, often forming several circular belts arranged on the lower part of stipe, a subapical to median, yellowish annulus often with an obvious brown edge, broadly ellipsoid to ellipsoid basidiospores (10.0–12.0 × 8.0–10.0 μm), and the presence of clamps. Furthermore, it is found in subalpine forests dominated by Picea and Abies.

In our multi-locus phylogenetic analysis (Fig. 1b), A. flavopantherina is, with low support, related to A. muscaria, A. regalis, A. persicina, A. ibotengutake, A. gemmata and A. concentrica. However, these species differ morphologically from A. flavopantherina in the following ways. Amanita muscaria, a species described from Europe and also found in northern, northeastern and northwestern China, has a red pileus and relatively narrower basidiospores measuring 9.0–12.5 × 7.0–8.5 μm (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015). The European A. regalis has a brown pileus covered with floccose to pulverulent volval remnants and relatively narrower basidiospores (9.0–11.0 × 7.0–8.0 μm) (Gilbert 1940, 1941a; Contu 2000a; Neville and Poumarat 2004). Amanita persicina has a pastel-red to light orange or melon-colored pileus, cream lamellae with a pinkish tint and relatively narrower basidiospores (9.4–12.7 × 7.0–8.5 μm) (Jenkins 1977, 1986; Tulloss et al. 2015). Amanita ibotengutake has white to grayish volval remnants on the pileus, smaller basidiospores (8.0–10.0 × 6.0–7.5 μm, Q = 1.23–1.38, Qm = 1.32 ± 0.06), and is found in temperate forests (Oda et al. 2002c; Yang 2015). Specimens of A. gemmata listed under A. concentrica differ from A. concentrica by their grayish yellow to yellowish pilei, volval remnants on the pileus composed primarily of subradically arranged elements with very abundant to nearly dominant filamentous hyphae, narrower basidiospores (9.5–12.0 × 6.5–8.5 μm), and absence of clamps. The comparisons between A. concentrica and A. flavopantherina are given in commentary on A. concentrica.

Amanita flavopantherina is easily confused with A. griseopantherina Yang-Yang Cui et al. as they have similar appearance and habitats, and for comparisons between the two species see the commentary on A. griseopantherina.

A species similar to A. flavopantherina and sharing a basidioma with clamps, also found in Abies forest is A. pakistanica Tulloss et al., but the latter differs by its umbonate pileus covered with patchy or flaky, white, detersile volval remnants, and relatively narrower basidiospores (10.1–12.2 × 7.0–8.8 μm) (Tulloss et al. 2001). Our phylogenetic analysis inferred from ITS data (Fig. S6) suggests that they represent two distinct species.

Amanita subglobosa Zhu L. Yang and A. pantherina (DC.) Krombh. are also similar to A. flavopantherina in their brown basidiomata. However, A. subglobosa has white volval remnants on the pileus and is dominant in forests of Pinaceae and Fagaceae at relatively lower altitudes (Yang 1997, 2005, 2015). Amanita pantherina has narrower basidiospores and no clamps (Gilbert 1940, 1941a; Yang 1997; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004). Molecular phylogenetic analyses (Figs. 1b, S6) indicate that they are distinct species.

6. Amanita griseopantherina Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 3c, d, 5.
Fig. 5

Microscopic features of Amanita griseopantherina (TYPE, HKAS 83560). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 824964

Etymology: griseopantherina, from griseus = gray, pantherina = Amanita pantherina, is proposed because this species is similar to the European A. pantherina, and has gray, pyramidal, verrucose to conical volval remnants on the pileus.

Type: CHINA. TIBET AUTONOMOUS REGION: Baxoi County, in a subalpine forest of Picea, altitude 3950 m, 29 June 2014, Qing Cai 1100 (TYPE, HKAS 83560, GenBank Acc. No.: ITS = MH508385, nrLSU = MH486573, tef1-α = MH508842, β-tubulin = MH485556).

Basidioma (Fig. 3c, d) medium-sized to large. Pileus 6–14 cm diam., convex, plano-convex to applanate, sometimes slightly depressed at center, yellow-brown (1B3–6), brownish (2C4–6), brown (3C2–6) to dark brown (4E4–7), often darker at center and becoming paler towards margin; volval remnants on pileus pyramidal, verrucose to conical, 1–6 mm diam., dirty white (2B1), white (1B1) to grayish (4B1–3), originally densely arranged over disk, but easily removed; margin slightly striate (ca. 0.1 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe 7–20 cm long × 1–3 cm diam., cylindric and slightly tapering upwards, with apex slightly expanded, dirty white (2B1) to white (1A1), sometimes with brownish (2C4–6) tinge, covered with white (1A1) to pale brownish (2B2–5) fibrils, often becoming floccose to patchy near basal bulb; context white (1A1), fistulose; basal bulb subglobose, fusiform to ellipsoid, 2–3.5 cm diam., white (1A1) to dirty white (2B1); volval remnants on stipe base collar-like, or shortly limbate often with its limb appressed to stipe, white (1A1), dirty white (2B1) to pale brownish (2B2–5). Annulus persistent, apical to subapical, white (1A1) to pale brownish (2B2–5), membranous. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–50 μm wide, composed of abundant elongate to subfusiform inflated cells (90–140 × 20–35 μm); filamentous hyphae fairly abundant to abundant, 3–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant clavate to long clavate inflated cells (60–110 × 15–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant, 2–7 μm wide. Subhymenium (Fig. 5a) 30–50 μm thick, with 2–3 layers of ovoid, subglobose to ellipsoid or irregular cells, 10–30 × 7–20 μm. Basidia (Fig. 5a) 50–60 (–70) × 12–15 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 5b) [100/7/7] (9.0–) 9.5–12.0 (–12.5) × (5.0–) 8.0–10.0 (–11.0) μm, Q = (1.1–) 1.15–1.31 (–1.44), Qm = 1.23 ± 0.07, mostly broadly ellipsoid, occasionally subglobose or ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, ovoid to ellipsoid or sphaeropedunculate inflated cells (14–35 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 120–300 μm thick; upper layer (70–150 μm thick) gelatinized, composed of radially, thin-walled, colorless or sometimes brownish to yellowish filamentous hyphae 2–7 μm wide; lower layer (50–80 μm thick) composed of radially and compactly arranged filamentous hyphae 3–10 μm wide, brownish to brown; vascular hyphae scarce. Volval remnants on pileus (Fig. 5c) composed of irregularly arranged elements: filamentous hyphae abundant, 3–7 (–9) μm wide, colorless or subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, subglobose, fusiform, ellipsoid to clavate, or sometimes irregular, 25–105 × 10–65 μm, colorless or subcolorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base dominantly composed of two parts intergrading to each other. Outer part dominantly composed of somewhat longitudinally arranged to interwoven elements: filamentous hyphae fairly abundant, 2–8 μm wide, colorless to subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to nearly dominant, becoming scarce and elongate towards stipitipellis, subglobose, fusiform, ellipsoid to clavate, 30–50 × 15–45 μm, colorless to subcolorless, terminal or in chains of 2–3; vascular hyphae scarce. Inner part dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 2–6 μm wide, colorless to subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to fairly abundant, fusiform, ellipsoid to clavate, 30–60 × 20–30 μm, colorless to subcolorless, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of outer part; inner surface similar to the structure of inner part but gelatinized and with more filamentous hyphae. Stipe trama composed of longitudinally arranged, clavate terminal cells, 80–300 × 15–45 μm; filamentous hyphae scattered to abundant, 2–13 μm wide; vascular hyphae scarce. Annulus dominantly composed of two parts intergrading to each other. Outer part dominantly composed of radially arranged to interwoven elements: filamentous hyphae very abundant to dominant, 3–6 μm wide, colorless to subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce and not easily observed; vascular hyphae scarce. Inner part non- to slightly gelatinized, composed of radially arranged elements: filamentous hyphae abundant to very abundant, 2–5 μm wide, colorless to subcolorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, elongate to clavate, 23–80 × 10–25 μm, colorless to subcolorless, terminal or in chains of 2–3; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in subalpine forests dominated by Abies and Picea; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. SICHUAN PROVINCE: Dege County, in a subalpine forest, altitude 3800 m, 9 August 2013, Xiao-Bin Liu 252 (HKAS 79953); Li County, in a subalpine forest with Abies and Picea, altitude 3170 m, 24 June 2014, Qing Cai 1086 (HKAS 83547); Tibetan Qiang Autonomous Prefecture of Ngawa, in a subalpine forest of Picea, altitude 3300 m, 12 August 2013, Bang Feng 1448 (HKAS 82340); Xiaojin County, in a subalpine forest, altitude 2330 m, 28 July 2007, Zai-Wei Ge 1602 (HKAS 53688). TIBET AUTONOMOUS REGION: Bomê County, in a subalpine forest of Abies, altitude 2763 m, 30 June 2014, Qing Cai 1118 (HKAS 83578). YUNNAN PROVINCE: Shangri-La, in a subalpine forest of Abies, altitude 3750 m, 25 August 2009, Qing Cai 136 (HKAS 58801).

Specimens of A. pantherina examined: CZECH REPUBLIC. CENTRAL BOHEMIAN REGION: 16 June 2009, J. Bororička s.n. (HKAS 56702). NETHERLANDS. SOUTH HOLLAND PROVINCE: Wassenaar, in a forest under Fagus, altitude unknown, 2 September 1979, C. Bas 7474 (L). GERMANY. HESSEN: Marburg, in a forest with Carpinus, Quercus and Tilia, altitude 250 m, 8 October 2004, KR 8678 (MB-102863).

Commentary: Amanita griseopantherina is characterized by its yellow-brown to brown pileus covered with pyramidal, verrucose to conical, white to dirty white volval remnants and with short striations on its margin, a white context, a white to dirty white stipe with collar-like or shortly limbate volva, an apical to subapical, white to pale brownish annulus, broadly ellipsoid basidiospores (9.5–12 × 8–10 μm), and the common presence of clamps. It is dominant in subalpine forests with Picea and Abies.

In our multi-locus phylogenetic analysis (Fig. 1b), A. griseopantherina is related to A. subglobosa, A. pantherina, A. pseudopantherina Zhu L. Yang et al. and A. velatipes G. F. Atk. However, A. subglobosa, a species described form southwestern China and also similar to A. griseopantherina in appearance, has a relatively smaller basidioma, and is found in forests dominated by Pinaceae and Fagaceae at relatively lower altitudes (Yang 1997, 2005, 2015). Amanita pantherina, a species described from Europe but not found in China, has a relatively lower annulus, narrower basidiospores and no clamps (Gilbert 1940, 1941a; Yang 1997; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; this study). Amanita pseudopantherina, a provisional name proposed by Yang (Yang 1997, 2005, 2015) and formally described as a novel species in this study, differs from A. griseopantherina by its relatively pale (brown to yellow-brown) pileus and the absence of clamps. Amanita velatipes, described from North America, can be easily distinguished from A. griseopantherina by its paler pileus covered with white patches, and smaller basidiospores (7.9–9.4 × 5.5–6.3 μm) (Murrill 1913; Gilbert 1941b; Jenkins 1977, 1982, 1986; Tulloss et al. 1995).

Amanita griseopantherina can be easily confused with A. flavopantherina in their similar appearance and habitats. However, A. flavopantherina has a darker brown pileus covered with yellow volval remnants, a white to yellow context, a relatively lower and yellower annulus with an obviously brown edge, a browner stipe, and broadly ellipsoid to ellipsoid basidiospores (10–12 × 8–10 μm).

Amanita pakistanica is similar to A. griseopantherina in the presence of clamps and Abies hosts. However, A. pakistanica has an umbonate pileus covered with patchy or flaky, detersile volval remnants and relatively narrower basidiospores (10.1–12.2 × 7–8.8 μm) (Tulloss et al. 2001). In our phylogenetic analysis inferred from ITS data (Fig. S6) they represent two distinct species.

Amanita ibotengutake, a species described from Japan and also distributed in northeastern China, is also similar to A. griseopantherina in their brown pileus and the common presence of clamps. However, A. ibotengutake has a relatively smaller and slender basidioma, and narrower basidiospores (8–10 × 6–7.5 μm) (Oda et al. 2002c; Yang 2015; see below).

7. Amanita ibotengutake T. Oda, C. Tanaka & Tsuda, Mycol. Prog. 1 (4): 360 (2002).

Pileus 7–9 cm in diam., yellow-brown (4B2–4) to brown (4C3–5); volval remnants on pileus pyramidal, verrucose to felted, white (1A1) to grayish (1B1–2), easily washed away; margin striate (0.1–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 7–13 cm long × 0.5–1.5 cm diam., cream (1A2) to white (1A1), with its upper part covered with pulverulent, white (1A1) squamules, lower part decorated with slightly recurved, white (1A1) to dirty white (2B1) squamules; context white (1A1), unchanging; basal bulb ovoid to subglobose, 1.5–2.5 cm diam.; volval remnants on stipe base conical to pulverulent, often formed an incomplete collar between lower part of stipe and stipe base. Annulus median.

Basidia 35–45 × 9–10 μm, clavate, 4-spored. Basidiospores [40/2/2] (7.5–) 8.0–10.0 × 6.0–7.5 μm, Q = (1.18–) 1.23–1.38 (–1.43), Qm = 1.32 ± 0.06, broadly ellipsoid, rarely ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged filamentous hyphae and inflated cells. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in temperate pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from northeastern China (Yang 2015; this study), Japan (Oda et al. 2002c) and Republic of Korea (Kim et al. 2013b).

Specimens examined: CHINA. JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest under Pinus koraiensis, altitude 680 m, 7 August 2010, Xiang-Hua Wang 2570 (HKAS 83269). LIAONING PROVINCE: Xifeng County, in a temperate forest, altitude 198 m, 2 August 2008, Yan-Chun Li 1191 (HKAS 56045).

Commentary: Amanita ibotengutake was described from Japan as a poisonous fungus because it produces the toxic substances ibotenic acid and muscimol (Oda et al. 2002c; Kim et al. 2013b). It was then reported from China and Republic of Korea (Kim et al. 2013b; Yang 2015).

Our multi-locus data (Fig. 1b) suggest that A. ibotengutake is likely to be related to A. muscaria, A. regalis, A. persicina, A. flavopantherina, A. gemmata and A. concentrica. However, these species differ from A. ibotengutake in the following ways. Amanita muscaria has a red pileus and broadly ellipsoid to ellipsoid basidiospores (9–12.5 × 7–8.5 μm) (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015). Amanita regalis has yellow volval remnants on the pileus and relatively broader basidiospores (9–11 × 7–8 μm) (Gilbert 1940, 1941a; Contu 2000a; Neville and Poumarat 2004). Amanita persicina has a pastel-red to light orange or melon-colored pileus covered with yellowish to tannish, floccose-fibrillose patches, cream lamellae with pinkish tint, a stipe whose context turns yellowish when cut or bruised and relatively larger basidiospores (9.4–12.7 × 7–8.5 μm) (Jenkins 1977, 1986; Tulloss et al. 2015).

For differences between A. ibotengutake and A. flavopantherina see the commentary on A. flavopantherina. Specimens of A. gemmata from Netherlands listed under A. concentrica differ from A. ibotengutake by their relatively longer basidiospores (9.5–12.0 × 6.5–8.5 μm) and the absence of clamps (this study).

The comparisons between A. ibotengutake and A. concentrica are given in the commentary on A. concentrica. For detailed descriptions, comparisons with A. subglobosa and images of A. ibotengutake see Yang (2015).

8. Amanita melleialba Zhu L. Yang, Qing Cai & Yang-Yang Cui, Fungal Divers. 15 (1): 189 (2015).

Pileus 3–5 cm in diam., honey yellow (2A6–8) to yellow (3A2–4) at center, becoming yellowish (2A2) to white (1A1) towards margin; volval remnants on pileus pyramidal to verrucose, ca. 1–2 mm in height, white (1A1) to yellowish (2A2); margin striate (0.5–0.6 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 4–8 cm long × 0.4–0.6 cm diam., white (1A1), covered with white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, 0.8–1.2 cm diam., with upper part covered with conical to pulverulent, white (1A1) to yellowish (2A2) volval remnants. Annulus subapical, white (1A1) with yellow (3A2–4) tinge.

Basidia 30–45 × 9–11 μm, clavate, 4-spored. Basidiospores [60/3/3] 7.5–9.5 (−10.0) × (5.5–) 6.0–7.0 μm, Q = (1.21–) 1.29–1.58 (−1.63), Qm = 1.41 ± 0.10, ellipsoid, sometimes broadly ellipsoid, inamyloid. Volval remnants on pileus composed of vertically to subvertically arranged elements: filamentous hyphae scarce to fairly abundant; inflated cells very abundant to nearly dominant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from central and southwestern China (Ariyawansa et al. 2015; Yang 2015).

Specimens examined: CHINA. HUNAN PROVINCE: Changsha, Yuelushan, in a subtropical forest, altitude ca. 260 m, 18 July 2014, S.C. Li 1 (HKAS 83216). YUNNAN PROVINCE: Puer, Caiyanghe, in a subtropical forest with Fagaceae, altitude 1300 m, 11 July 2014, Gang Wu 1339 [HOLOTYPE, HKAS 83446, as Amanita sp. 1 in Yang (2015)]; same location, in a subtropical forest with Fagaceae, altitude 1326 m, 11 July 2014, Xiao-Bin Liu 439 (HKAS 87085).

Commentary: This species corresponds to “Amanita sp. 1” in Yang (2015) and was formally described from China by Ariyawansa et al. (2015). Its phylogenetic related species were yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. melleialba see Ariyawansa et al. (2015) and Yang (2015).

9. Amanita melleiceps Hongo, J. Jpn. Bot. 41: 165 (1966).

Basidioma small to medium-sized. Pileus 2–5 cm in diam., yellow (3A2–4) to honey yellow (2A6–8) at center, but becoming white (1A1) toward margin; volval remnants on pileus patchy to felted, yellowish (1A2) to dirty white (1A1); margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 3–7 cm long × 0.3–0.6 cm diam., white (1A1) to cream (1A2); context white (1A1), unchanging; basal bulb ovoid to subglobose, 0.5–1 cm diam., with its upper half covered with pulverulent, white (1A1) to yellowish (1A2) volval remnants. Annulus absent.

Basidia 40–55 × 8–11 μm, clavate, 4-spored. Basidiospores [160/8/4] (7.5–) 8.5–10.5 (–11.5) × (5.5–) 6.0–7.5 (–8.0) μm, Q = (1.21–) 1.27–1.58 (–1.77), Qm = 1.42 ± 0.1, broadly ellipsoid to ellipsoid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae fairly abundant; inflated cells abundant to very abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine (e.g. Pinus massoniana) or mixed (Fagaceae and Pinaceae) forests; basidioma occurring in summer and autumn.

Distribution: Known from central and southern China (Yang 2005, 2015; this study), Japan (Hongo 1966; Yang and Doi 1999; Imazeki et al. 2011) and Republic of Korea (Kim et al. 2013b).

Specimens examined: CHINA. ANHUI PROVINCE: Qianshan County, in a forest of Pinus massoniana, altitude 120 m, 28 June 2013, Qi Zhao 1830 (HKAS 80940). HENAN PROVINCE: Shangcheng County, in a forest with Fagaceae and Pinus massoniana, altitude 200 m, 29 June 2013, Yan-Jia Hao 904 (HKAS 80184). HUBEI PROVINCE: Macheng, Wunaoshan, in a forest with Fagaceae and Pinaceae, altitude 100 m, 26 June 2013, Yan-Jia Hao 865 (HKAS 80145). JAPAN. HONSHU: Senjô, in a pine forest mixed with Quercus, altitude unknown, 22 June 1965, Hongo Herb. 3070 (HOLOTYPE, TNS-F-237279).

Commentary: Amanita melleiceps was described from Japan by Hongo (1966). It was subsequently reported from China and the Republic of Korea (Yang 2005, 2015; Kim et al. 2013b).

Our multi-locus phylogenetic analysis (Fig. 1b) suggests that A. melleiceps is related to A. breckonii Thiers & Ammirati. However, the latter differs by its relatively larger and robuster basidioma with a pileus ca. 4–9 cm in diam., volval remnants on the stipe base forming a low rim around the apex of basal bulb, and relatively larger basidiospores (9.6–12.0 × 5.6–8.0 μm) (Thiers 1982; Thiers and Ammirati 1982; Jenkins 1986). For detailed descriptions, comparisons with similar species, line drawings and images of A. melleiceps see Yang (2005, 2015).

10. Amanita mira Corner & Bas, Persoonia 2(3): 290 (1962).

Pileus 4–8 cm in diam., yellow (3A4–7) to yellow-brown (3B3–7); volval remnants on pileus pyramidal to conical, cream, yellowish (3A2–4) to yellow (4A4–6), removable; margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 5–8 cm long × 0.8–1.2 cm diam., white (1A1) to cream (1A2); context white (1A1), unchanging; basal bulb ovoid to fusiform, 1–2 cm diam., with upper part covered with verrucose, floccose to pulverulent, yellow (4A4–6) volval remnants, often arranged in incomplete rings. Annulus absent.

Basidia 30–45 × 8–12 μm, clavate, 4-spored. Basidiospores [125/5/3] (6–) 6.5–8.0 (–9.0) × (5.0–) 6.0–7.5 (–8.0) μm, Q = 1.0–1.15 (–1.21), Qm = 1.07 ± 0.04, often globose to subglobose, rarely broadly ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae scarce; inflated cells very abundant. Volval remnants on stipe base composed of irregularly to regularly arranged elements: filamentous hyphae very abundant; inflated cells scarce, but abundant on the apical part of verrucose volval remnants. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests with Fagaceae (e.g. Castanopsis and Lithocarpus); basidioma occurring in summer.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study), Malaysia (Lee 2017) and Singapore (Corner and Bas 1962; Lee 2017).

Specimens examined: CHINA. YUNNAN PROVINCE: Mengla County, in a subtropical broad-leaved forest, altitude 1000 m, 12 August 1995, Zhu L. Yang 2163 (HKAS 29526); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 1000 m, 4 July 2015, Yang-Yang Cui 221 (HKAS 92122); same location, in a mixed forest with Lithocarpus and Pinus, altitude 800 m, 4 July 2015, Qing Cai 1330 (HKAS 91953).

Commentary: Amanita mira was described from Singapore (Corner and Bas 1962), and has been collected in Malaysia and tropical to subtropical regions of China with broad-leaved trees (Yang 1997, 2005, 2015; Lee 2017).

Based on our multi-locus phylogenetic study (Fig. 1b), A. mira is related to A. elata, A. sychnopyramis f. sychnopyramis and A. sychnopyramis f. subannulata. However, A. elata has a pileus yellowish brown at center but yellowish to white at margin, floccose to pulverulent volval remnants on pileus and relatively larger basidiospores (7.5–9.5 × 7.0–9.0 μm) (Corner and Bas 1962; Pegler 1986; Adhikari 1999; Bhatt et al. 2003; Pradeep and Vrinda 2010; Yang 2015; Lee 2017). Amanita sychnopyramis f. sychnopyramis and A. sychnopyramis f. subannulata have brownish, gray-brown to dark brown pilei and dirty white to grayish volval remnants on the pileus (Corner and Bas 1962; Hongo 1971; Yang 1997, 2005, 2015; Lee 2017). Furthermore, A. sychnopyramis f. subannulata has a median annulus (Hongo 1971; Yang 1997, 2005, 2015).

For detailed descriptions, comparisons with similar species, line drawings and images of A. mira see Yang (1997, 2005, 2015).

11. Amanita muscaria (L.: Fr.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 111 (1783).

Basionym: Agaricus muscarius L., Sp. pl. 2: 1172 (1753).

Pileus 5–15 cm in diam., red (7A7–8) to orange-red (6A5–8), sometimes with yellow (4A4) tinge; volval remnants on pileus pyramidal, conical, verrucose to felted, white (1A1) to dirty white (2B1), sometimes yellowish (2A2), removable; margin striate (up to 0.2 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 7–18 cm long × 0.5–2.5 cm diam., white (1A1), covered with white (1A1) fibrils; context white (1A1), unchanging; basal bulb ovoid, fusiform to subglobose, 1–4 cm diam.; volval remnants on stipe base verrucose to conical, white (1A1) to yellowish (2A2), sometimes arranged in incomplete rings. Annulus subapical to submedian.

Basidia 40–60 × 12–15 μm, clavate, 4-spored. Basidiospores [70/6/6] 9.0–12.5 × 7.0–8.5 μm, Q = 1.24–1.47, Qm = 1.34 ± 0.08, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae scattered; inflated cells very abundant. Volval remnants on stipe base similar to structure of volval remnants on pileus, but with irregularly arranged elements. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in temperate, coniferous or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from Asia (Imai 1933, 1938; Kumar et al. 1990; Yang 2005, 2015; Geml et al. 2006, 2008; Imazeki et al. 2011), Europe (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004) and North America (Coker 1917; Thiers 1982; Jenkins 1977, 1986; Geml et al. 2006, 2008); in China only known from northeastern and northwestern areas.

Specimens examined: CHINA. HEILONGJIANG PROVINCE: Daxinganling, in a temperate forest, altitude 1200 m, 21 August 2010, Xiang-Hua Wang 2691 (HKAS 61888); Yitulihegoukou, in a temperate forest, altitude unknown, 16 August 1974, “Heilongjiang Biologic Institute” 9 (HMAS 37367). XINJIANG UYGUR AUTONOMOUS REGION: Aertaishan, altitude unknown, August 1996, Jun-Yan Wang 490 (HMAS 63490). AUSTRIA. VORARLBERG: Kleinwalsertal, in a forest with Abies and Picea, altitude 1200 m, 29 September 2016, Zhu L. Yang 5966 (HKAS 96165). RUSSIA: Kyrov, Kofelnych, altitude unknown, 14 August 2016, collector unknown (HMJAU 4549); Kyrov, Orlovsky, altitude unknown, 11 August 2006, collector unknown (HMJAU 4585).

Commentary: Amanita muscaria is well known as fly agaric and has been traditionally used to catch flies, because the psychoactive substances including muscarine and isoxazole derivatives it contains supposedly attract insects (Crundwell 1987; Michelot and Melendez-Howell 2003; Satora et al. 2005; Chen et al. 2016; Lumpert and Kreft 2016; Mikaszewska-Sokolewicz et al. 2016). This species is described from Europe and represents the type species of the genus Amanita (Gilbert 1940, 1941a; Jenkins and Petersen 1976; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015). Yang and Oberwinkler (1999) provided a detailed study on the basidiomal development and anatomy of A. muscaria.

Amanita muscaria was traditionally treated as a single morphospecies with several varieties (Jenkins 1977, 1986; Thiers 1982; Contu 2000a; Wasser 2000; Neville and Poumarat 2004; Geml et al. 2006, 2008) and has been reported as a common, widespread, easily identifiable, ecologically plastic fungus with a wide niche (Coker 1917; Jenkins 1977, 1986; Reid 1980; Thiers 1982; Garrido 1986; Reid and Eicker 1991; Ridley 1991; Tulloss et al. 1995; Bhatt et al. 2003; Oda et al. 2004; Yang 2005, 2015; Vargas et al. 2017). However, the studies of Geml et al. (2006) and Geml et al. (2008) indicated that the morphospecies A. muscaria represents more than one phylogenetic species.

In the multi-locus analysis (ITS + β-tubulin + LSU + ef1-α) of Geml et al. (2008), the morphospecies A. muscaria was divided into eight clades with strong inter- and intracontinental phylogeographic structure. In our ITS analysis (Fig. S6), our Chinese specimens, HMJAU 4548, HMJAU 4549, and HKAS 61888, clustered within the clade II of Geml et al. (2008), which contained many specimens from Eurasia and Alaska.

In our multi-locus analysis (Fig. 1b), A. muscaria was found to be closely related to A. persicina. However, A. persicina has a relatively paler colored (pastel-red to light orange or melon-colored) pileus covered with pale yellow to yellowish-tan to tan, floccose-fibrillose patches and cream lamellae with a pinkish tint (Jenkins 1977, 1986; Tulloss et al. 2015).

12. Amanita orientigemmata Zhu L. Yang & Yoshim. Doi, Bull. natn. Sci. Mus., Tokyo, Ser. B 25(3): 107 (1999).

Synonym: Amanita pyriformis Boonprat. & Parnmen, Fungal Diversity 78: 138 (2016), syn. nov.

Pileus 4–10 cm in diam., yellow (2B2–5) to yellowish (2A2–4); volval remnants on pileus felted to patchy, white (1A1) to dirty white (2B1), removable; margin striate (0.1–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 6–12 cm long × 0.5–1 cm diam., white (1A1) to cream (1A2); context white (1A1), unchanging; basal bulb subglobose, 1–2 cm diam., with upper part covered with patchy, white (1A1) to yellowish (2A2–4) volval remnants. Annulus white (1A1), easily broken and fugacious.

Basidia 38–60 × 10–14 μm, clavate, 4-spored. Basidiospores [40/6/6] (7.0–) 8.0–10.0 (–10.5) × (5.5–) 6.0–7.5 μm, Q = (1.07–) 1.13–1.51 (–1.55), Qm = 1.3 ± 0.14, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae fairly abundant; inflated cells fairly abundant to abundant. Volval remnants on stipe base similar to structure of volval remnants on pileus, but with irregularly arranged elements. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer.

Distribution: Known from northeastern, northwestern and southern China (Yang 2005, 2015; this study), India (Bhatt et al. 2003) and Japan (Yang and Doi 1999).

Specimens examined: CHINA. FUJIAN PROVINCE: Wuyishan, in a forest of Pinus massoniana, altitude 700–800 m, 6 August 2001, Yan Liu 010806023 (HKAS 38812). JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest, altitude ca. 1000 m, date unknown, Yun Wang 811333 (IFP). SHAANXI PROVINCE: Zhenping County, in a forest with Quercus, altitude 1200 m, 4 July 2013, Qi Zhao 1868 (HKAS 80978). JAPAN. HONSHU: Tottori, 12 September 1979, E. & F. Nagasawa s.n. (HOLOTYPE, TNS-F-198537); same location, 8 September 1979, E. & F. Nagasawa s.n. (TNS-F-198540); same location, 9 September 1979, E. & F. Nagasawa s.n. (TNS-F-198546).

Commentary: Amanita orientigemmata was described from Japan by Yang and Doi (1999). It was subsequently reported from both China and India (Bhatt et al. 2003; Yang 2005, 2015). It is characterized by its yellowish to yellow pileus covered with white to dirty white, felted, patchy or subpyramidal volval remnants, white to yellowish, patchy to verrucose or sometimes appressed-limbate volval remnants on the stipe base, broadly ellipsoid to ellipsoid basidiospores and the common presence of clamps.

In our ITS phylogenetic analysis, A. pyriformis clustered within A. orientigemmata as a monophyly with strong statistical support (Fig. S6). We therefore treat A. pyriformis as a synonym of A. orientigemmata. The presence of cystidia reported in A. pyriformis (Li et al. 2016) should a misinterpretation.

For detailed descriptions, comparisons with similar species, line drawings and images of A. orientigemmata see Yang and Doi (1999), and Yang (2005, 2015).

13. Amanita parvipantherina Zhu L. Yang, M. Weiß & Oberw., Mycologia 96(3): 639 (2004).

Basidioma small to medium-sized. Pileus 3–6 cm in diam., brown (2B2–3, 3C2–4); volval remnants on pileus verrucose to pyramidal, 1–2 mm in height, cream (1A2), white (1A1) to dirty white (2B1); margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 4–10 cm long × 0.5–1 cm diam., yellowish (1A3–5), cream (1A2) to white (1A1); context white (1A1), unchanging; basal bulb ovoid to subglobose, 1–2 cm diam., with its upper part covered with floccose to granular, white (1A1), cream (1A2) to brownish (2B2–3) volval remnants. Annulus subapical, white (1A1) to brownish (2B2–3).

Basidia 45–58 × 11–14 μm, clavate, 4-spored. Basidiospores [360/17/16] (8.0–) 8.5–11.5 (–13.5) × (6.0–) 7.0–8.5 (–9.0) μm, Q = 1.13–1.47, Qm = 1.29 ± 0.11, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae fairly abundant; inflated cells abundant to nearly dominant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang et al. 2004; Yang 2005, 2015; this study), and India (Bhatt et al. 2017).

Specimens examined: CHINA. YUNNAN PROVINCE: Baoshan, in a forest with Pinus yunnanensis, altitude 1900 m, 10 August 2010, Qing Cai 343 (HKAS 67907); same location, in a pine forest, altitude ca. 1900 m, 9 August 2010, Qing Cai 330 (HKAS 67892); same location, in a mixed forest with Fagaceae and Pinaceae, altitude ca. 2000 m, 12 August 2011, Yan-Jia Hao 493 (HKAS 71602); Changning County, in a mixed forest with Fagaceae and Pinaceae, altitude 2016 m, 25 July 2009, Li-Ping Tang 935 (HKAS 56892); Fengqing County, in a mixed forest with Fagaceae and Pinaceae, altitude 1979 m, 26 July 2009, Li-Ping Tang 963 (HKAS 56920); Fugong County, in a pine forest, altitude 1200 m, 4 August 2010, Qing Cai 266 (HKAS 67828); Heqing County, Huangping, in a forest of Pinus yunnanensis, altitude 2500 m, 24 July 1998, Zhu L. Yang 2490 (HOLOTYPE, HKAS 32492); Kunming, Kunming Institute of Botany, altitude 1980 m, 25 August 2008, Zhu L. Yang 5140 (HKAS 54437); Mengla County, in a mixed forest with Fagaceae and Pinaceae, altitude 1200 m, 9 July 2014, Gang Wu 1296 (HKAS 89033); Nanjian County, Lingbaoshan, in a broad-leaved forest of Castanopsis and Lithocarpus, altitude 2350 m, 3 August 2014, Qing Cai 1203 (HKAS 83663); same location, in a broad-leaved forest of Castanopsis and Lithocarpus, altitude ca. 2300 m, 3 August 2014, Qing Cai 1194 (HKAS 83654); same location, in a broad-leaved forest with Castanopsis and Lithocarpus, altitude ca. 2300 m, 3 August 2014; Qing Cai 1206 (HKAS 83673); Longyang, in a mixed forest with Fagaceae and Pinaceae, altitude 1761 m, 23 July 2009, Li-Ping Tang 897 (HKAS 56854); Simao, in a subtropical forest, altitude 1600 m, 30 July 2008, Li-Ping Tang 519 (HKAS 54750); Simao, Caiyanghe, in a subtropical forest dominated by Fagaceae, altitude 1400 m, 28 July 2008, Li-Ping Tang 492 (HKAS 54723); Tengchong County, in a forest of Pinus armandii and Keteleeria fortunei, altitude 2010 m, 14 August 2010, Qing Cai 400 (HKAS 67962).

Commentary: Amanita parvipantherina was described from China by Yang et al. (2004), and subsequently reported from India (Bhatt et al. 2017). It is characterized by its small to medium-sized basidioma, a brown pileus covered with cream, white to dirty white, verrucose to pyramidal volval remnants, a subapical annulus, broadly ellipsoid to ellipsoid basidiospores and the absence of clamps. The species to which it is closely phylogenetically related are as yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species, line drawings and images of A. parvipantherina see Yang et al. (2004) and Yang (2005, 2015).

14. Amanita pseudopantherina Zhu L. Yang ex Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov. [Amanita pseudopantherina Zhu L. Yang, Bibl. Mycol. 170: 44 (1997), nom. prov., inval.]

Figures 3e, 6.
Fig. 6

Microscopic features of Amanita pseudopantherina (TYPE, HKAS 80007). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 824969

Etymology: pseudopantherina, from pseudo- = false-, and pantherina = Amanita pantherina, is proposed because this species is similar to European A. pantherina.

Type: CHINA. YUNNAN PROVINCE: Binchuan County, Jizushan, in a forest with Fagaceae, altitude 2300 m, 4 August 2013, Zhu L. Yang 5727 (TYPE, HKAS 80007, GenBank Acc. No.: ITS = MH508514, nrLSU = MH486777, tef1-α = MH509004, rpb2 = MH486191, β-tubulin = MH485698).

Basidioma (Fig. 3e) medium-sized to large. Pileus 3.5–12 cm diam., plano-convex to applanate, lacking a depression or umbo at center, brownish (1B2–5), yellow-brown (2B2–5) to brown (3C2–5); volval remnants on pileus pyramidal, verrucose to subconical, dirty white (2B1) to white (1A1); margin slightly striate (ca. 0.25–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe 6.5–16 cm long × 0.7–1.5 cm diam., cylindrical and slightly tapering upwards, with apex slightly expanded, dirty white (2B1) to white (1A1), sometimes with yellowish (2A2–5) tinge, glabrous or covered with concolorous, floccose squamules; context white (1A1), hollow in center; basal bulb globose to subglobose, 1–3 cm diam., white (1A1) to dirty white (2B1); volval remnants on stipe base floccose, arranged in belts on lower part of stipe, and often formed a collar-like or shortly limbate volva on limit between stipe and basal bulb, white (1A1) to dirty white (2B1), sometimes with yellowish (2A3–5) tinge. Annulus persistent, subapical, pendant from attachment ca. 2–4 cm below apex of stipe, white (1A1), membranous, with a brownish (2B2–5) to brown (3C2–5) edge. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant ellipsoid inflated cells (50–80 × 15–25 μm); filamentous hyphae fairly abundant to abundant, 3–6 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid inflated cells (50–80 × 10–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant, 3–5 μm wide. Subhymenium (Fig. 6a) 30–60 μm thick, with 2–3 layers of subglobose, fusiform, ellipsoid or irregular cells, 10–30 × 8–25 μm. Basidia (Fig. 6a) 40–50 × 10–14 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 6b) [100/8/7] (9.0–) 9.5–12.0 (–12.5) × (5.0–) 8.0–10.0 (–11.0) μm, Q = 1.1–1.36 (–1.44), Qm = 1.23 ± 0.07, mostly broadly ellipsoid, sometimes subglobose or ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid inflated cells (10–30 × 8–12 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–5 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 80–200 μm thick; upper layer (35–75 μm thick) gelatinized, composed of radially, thin-walled, colorless filamentous hyphae 2–8 μm wide; lower layer (75–100 μm thick) composed of radially and compactly arranged, filamentous hyphae 2–7 μm wide, yellow or yellow-brown; vascular hyphae scarce. Volval remnants on pileus (Fig. 6c) composed of vertically arranged elements: filamentous hyphae scarce to fairly abundant, 2–9 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to dominant, globose, subglobose, fusiform to ellipsoid, 25–70 × 15–40 μm, colorless, thin-walled (≤ 1 μm), terminal or in chains of 2–3; vascular hyphae scarce. Interior of volval remnants on stipe base dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant, 3–7 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, fusiform to ellipsoid, 40–70 × 25–50 μm, colorless, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of interior part, but with scattered to fairly abundant inflated cells; inner surface similar to the structure of interior part, but gelatinized. Stipe trama composed of longitudinally arranged, clavate terminal cells, 170–480 × 20–35 μm; filamentous hyphae scattered to abundant, 3–12 μm wide; vascular hyphae scarce. Annulus dominantly composed of radially arranged elements: filamentous hyphae very abundant, 3–7 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to fairly abundant, subglobose, fusiform to elongate, 20–60 × 10–35 μm, colorless, terminal or in chains of 2–3; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. SICHUAN PROVINCE: Tibetan Autonomous County of Muli, in a subalpine forest of Picea, altitude 3600 m, 1 August 2012, Ting Guo 536 (HKAS 76228). YUNNAN PROVINCE: Binchuan County, Jizushan, in a forest with Fagaceae, altitude 1750 m, 8 August 2014, Qing Cai 1278 (HKAS 83747); Diqing, Habaxueshan, in a mixed forest, altitude 3100 m, 14 August 2008, Li-Ping Tang 638 (HKAS 54869); Lanping County, in a forest of Pinus yunnanensis, altitude 2510 m, 13 August 2010, Xue-Tai Zhu 138 (HKAS 68314); Nanjian County, Wuliangshan, in a forest dominated by Fagaceae, altitude 2229 m, 22 August 2014, Qing Cai 1176 (HKAS 83636); Yongping County, in a forest with Fagaceae and Pinaceae, altitude 2400 m, 1 August 2009, Gang Wu 79 (HKAS 57611).

Commentary: Amanita pseudopantherina is characterized by its brown pileus densely covered with white, pyramidal volval remnants, a globose to subglobose stipe base covered with collar-like, limbate volval remnants, and broadly ellipsoid basidiospores (9.5–12.0 × 8.0–10.0 μm).

In our ITS phylogenetic analysis (Fig. S6), A. pseudopantherina was closely related to A. pantherina. However, the European A. pantherina has a robust basidioma, and narrower basidiospores (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; this study).

In China, species in Amanita sect. Amanita with brown pileus covered with white, pyramidal, verrucose, conical volval remnants, and shortly limbate volval remnants on the stipe base comparable to A. pseudopantherina include A. parvipantherina, A. subglobosa, and A. subparvipantherina Zhu L. Yang et al. However, A. parvipantherina has a relatively smaller and slender basidioma, and relatively narrower basidiospores (8.5–11.5 × 6.5–8.5 μm) (Yang et al. 2004; Yang 2005, 2015). Amanita subglobosa can be distinguished from A. pseudopantherina by its relatively darker colored pileus and the common presence of clamps (Yang 1997, 2005, 2015). Amanita subparvipantherina possesses narrower basidiospores (9.0–11.5 × 6.5–8.0 μm) and more abundant inflated cells in the volval remnants on the pileus (Ariyawansa et al. 2015; see below).

The North American A. velatipes is somewhat similar to A. pseudopantherina, but the former has a paler yellow pileus covered with warted to patchy volval remnants, and relatively smaller basidiospores (7.9–9.4 × 5.5–6.3 μm) (Murrill 1913; Gilbert 1941b; Jenkins 1977, 1982, 1986; Tulloss et al. 1995).

15. Amanita pseudosychnopyramis Yang-Yang Cui, Qing Cai & Zhu L. Yang, Fungal Divers. 15 (1): 190 (2015).

Pileus 4–7 cm in diam., yellowish brown (3B2–8) to brownish (3C4–7) at center, becoming yellowish (3A2–3, 3B2–4) to dirty white (3B1–2) towards margin; volval remnants on pileus conical, subconical to pyramidal, dirty white, grayish (2B1) to brownish gray (3C2–3); margin striate (0.25–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae truncate. Stipe 6–8.5 cm long × 0.3–0.8 cm diam., dirty white (2B1) to yellowish (3A2–3, 3B2–4); context white (1A1), unchanging; basal bulb subglobose to long ellipsoid, 1–1.3 cm diam., with upper part covered with conical, subconical to granular, grayish (2C1) to brownish gray (3C2–3) volval remnants arranged in several incomplete rings, usually forming a short limbate collar between stipe and bulb. Annulus subapical.

Basidia 36–47 × 10–13 μm, clavate, 4-spored, sometimes 2-spored. Basidiospores [140/7/3] (7.0–) 8.0–10.0 (–11.5) × (6.5–) 7.0–8.5 (–10.5) μm, Q = 1.06–1.20 (–1.39), Qm = 1.13 ± 0.05, subglobose to broadly ellipsoid, inamyloid. Volval remnants on pileus composed of more or less vertically arranged elements: filamentous hyphae abundant; inflated cells very abundant to nearly dominant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical forests dominated by Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China (Ariyawansa et al. 2015; this study).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 500 m, 20 March 2012, Fang Li 66 (HKAS 78417); same location, in a subtropical forest dominated by Fagaceae, altitude 320 m, 16 March 2014, Fang Li 1593 (HKAS 82293). YUNNAN PROVINCE: Jinghong, Dadugang, in a subtropical forest dominated by Fagaceae, altitude ca. 1000 m, 30 June 2014, Kuan Zhao 467 (HOLOTYPE, HKAS 87999).

Commentary: Amanita pseudosychnopyramis was described from China by Ariyawansa et al. (2015). This species can be recognized by its conical to pyramidal, gray to brownish gray volval remnants on the pileus, a basal bulb with a short limbate volva on the stipe base and relatively larger basidiospores. Species with which it is phylogenetically related are as yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. pseudosychnopyramis see Ariyawansa et al. (2015).

16. Amanita rubrovolvata S. Imai, Bot. Mag. (Tokyo) 53: 392 (1939).

Pileus 2–6.5 cm in diam., red (6A6–8) to orange-red (5A6–8) at center, but becoming orange (5A3–6) to yellow (4A2–3) towards margin; volval remnants on pileus pulverulent to conical, red (6A6–8), orange (5A3–6) to yellow (4A2–3); margin striate (0.2–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 5–10 cm long × 0.5–1 cm diam., cream (1A2) above annulus, cream (1A2) to yellowish (1A3–5) below annulus; context white (1A1), unchanging; basal bulb ovoid to subglobose, 1–2 cm diam., with its upper part covered with white (1A1), cream (1A2) to brownish (3C2) volval remnants. Annulus subapical, white (1A1) to brownish (3C2).

Basidia 35–48 × 9–11 (–13) μm, clavate, 4-spored. Basidiospores [280/14/11] (7.0–) 7.5–9.0 (–11.0) × (6.5–) 7.0–8.5 (–10.5) μm, Q = 1.0–1.1 (–1.23), Qm = 1.06 ± 0.04, globose to subglobose, inamyloid. Volval remnants on pileus composed of vertically to irregularly arranged elements: filamentous hyphae scarce to scattered; inflated cells abundant. Volval remnants on stipe base similar to structure of volval remnants on pileus, but with more filamentous hyphae. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, eastern, southern and southwestern China (Yang 1997, 2005, 2015; this study), Malaysia (Corner and Bas 1962), India (Bhatt et al. 2003; Semwal et al. 2007), Japan (Imai 1939; Imazeki et al. 2011), Republic of Korea (Kim et al. 2013b) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. HUNAN PROVINCE: Yanling County, in a subtropical forest with Fagaceae, altitude 1516 m, 22 July 2012, Qing Cai 865 (HKAS 75599). YUNNAN PROVINCE: Baoshan, Longyang, in a mixed forest with Fagaceae and Pinaceae, 1900 m, 13 August 2011, Gang Wu 658 (HKAS 81993); Binchuan County, Jizushan, in a subtropical forest of broad-leaved trees, altitude 2200 m, 4 August 2013, Zhu L. Yang 5725 (HKAS 80006); Lanping County, in a forest with Pinus yunnanensis, altitude 2400 m, 14 August 2010, Xue-Tai Zhu 146 (HKAS 68322); Lijiang Alpine Botanic Garden, in a forest of Quercus, altitude 3500 m, 8 August 2014, Xiao-Bin Liu 506 (HKAS 87151); Lushui County, in a mixed forest, altitude 2000 m, 7 August 2010, Li-Ping Tang 1193 (HKAS 63011); Puer, altitude 1700 m, 7 September 2008, Zhu L. Yang 5194 (HKAS 54491); Simao, Caiyanghe, in a subtropical broad-leaved forest, altitude 1400 m, 28 July 2008, Li-Ping Tang 494 (HKAS 54725); Yingjiang County, in a subtropical broad-leaved forest with Fagaceae, altitude 1420 m, 16 July 2009, Li-Ping Tang 787 (HKAS 56744). GUANGXI PROVINCE: Xingan County, Maoershan, in a broad-leaved forest, altitude 1900 m, 24 July 2012, Qing Cai 877 (HKAS 75611). TAIWAN PROVINCE: Nantou County, Hehuanshan, in a forest of Abies, altitude 3100 m, 15 September 2012, Bang Feng 1265 (HKAS 82407).

Commentary: Amanita rubrovolvata was described from Japan by Imai (1939), and subsequently reported from China, Malaysia, India, the Republic of Korea and Thailand (Corner and Bas 1962; Yang 1997, 2005, 2015; Bhatt et al. 2003; Semwal et al. 2007; Sanmee et al. 2008; Kim et al. 2013b). This species can be easily recognized by its small red pileus and pulverulent, red to orange volval remnants on the stipe base. Species with which it is phylogenetically closely related are as yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species, line drawings and images of A. rubrovolvata see Yang (1997, 2005, 2015).

17. Amanita rufoferruginea Hongo, J. Jpn. Bot. 41: 165 (1966).

Pileus 4–7 cm in diam., yellow-brown (4B5–7) to orange-brown (5B4–7), densely covered with pulverulent, concolorous volval remnants; margin striate (0.2–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 7–10 cm long × 0.5–1 cm diam., densely covered with pulverulent, red (5A7) to yellow-brown (4B5–7) squamules; context white (1A1), unchanging; stipe base subglobose, 1.5–2 cm diam., with its upper part covered with verrucose to pulverulent, orange-red (5A5–8) to yellow-brown (4B5–7) volval remnants. Annulus apical, fragile and fugacious.

Basidia 30–48 × 10–14 μm, clavate, 4-spored. Basidiospores [90/12/12] 7.0–9.0 (–13) × 6.5–8.5 (–11.5) μm, Q = (1.0–) 1.04–1.16 (–1.23), Qm = 1.09 ± 0.04, subglobose, sometimes globose, inamyloid. Volval remnants on pileus composed of irregularly, loosely arranged elements: filamentous hyphae scarce to fairly abundant; inflated cells very abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, eastern, southern and southwestern China (Yang 2005, 2015; this study), Japan (Hongo 1966, 1967; Imazeki et al. 2011) and Republic of Korea (Park et al. 1990).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 850 m, 3 June 2013, Qing Cai 937 (HKAS 79616); same location, in a subtropical forest dominated by Fagaceae, altitude ca. 800 m, 1 June 2013, Qing Cai 913 (HKAS 79592); same location, in a subtropical forest dominated by Fagaceae, altitude ca. 800 m, 9 May 2012, Fang Li 196 (HKAS 78421); same location, in a forest dominated by Fagaceae, altitude ca. 540 m, 10 May 2012, Fang Li 214 (HKAS 78428); same location, in a subtropical forest dominated by Fagaceae, altitude 500 m, date unknown, Fang Li 916 (HKAS 101395); same location, in a subtropical forest dominated by Fagaceae, altitude 550 m, 17 May 2012, Fang Li 311 (HKAS 101396). GUANGXI PROVINCE: Qinglong, altitude unknown, 24 May 1970, Yu-Chen Zong et al. 58 (HMAS 35979). GUIZHOU PROVINCE: altitude unknown, August 1983, Xing-Liang Wu 817 (HKAS 14378). HUNAN PROVINCE: Changsha, in a subtropical forest with Pinus and Lithocarpus, altitude ca. 100 m, 11 June 2001, Zhu L. Yang 3045 (HKAS 38123). TAIWAN PROVINCE: Nantou County, altitude unknown, 25 October 2001, Jian-Ming Chen 3099 (HKAS 38787). YUNNAN PROVINCE: Puer, in a subtropical forest, altitude ca. 1600 m, date unknown, Shu Yao 6 (HKAS 84974). JAPAN. KYOTO: Iwakura, in a pine forest, altitude unknown, 29 July 1965, Hongo Herb. 3122 (HOLOTYPE, TNS-F-244373).

Commentary: Amanita rufoferruginea was described from Japan, and then reported from China and the Republic of Korea (Hongo 1966, 1967; Park et al. 1990; Yang 2005, 2015; Imazeki et al. 2011). This species can be easily recognized by its yellow-brown basidioma densely covered with pulverulent, orange-red to yellow-brown volval remnants, and subglobose basidiospores (Hongo 1966, 1967; Imazeki and Hongo 1987; Yang 1997, 2005, 2015). Phylogenetically, A. rufoferruginea is closely related to A. siamensis Sanmee et al. (Fig. 1b). Indeed, they are also very similar to each other morphologically. For comparisons between the two species see the commentary on A. rufoferruginea in Yang (2015).

18. Amanita siamensis Sanmee, Zhu L. Yang, P. Lumyong & Lumyong, Mycotaxon 88: 225 (2003).

Pileus 5–7 cm in diam., yellowish brown (5C2–6), with olivaceous (30A6–8) tinge, densely covered with pulverulent, yellow-brown (5C2–6) volval remnants; margin striate (0.2–0.4 R), non-appendiculate. Lamellae white (1A1), lamellulae truncate. Stipe 7–10 cm long × 0.7–1 cm diam., densely covered with pulverulent, yellow-brown (5C2–6) squamules, with olivaceous (30A6–8) tinge; basal bulb fusiform to ellipsoid, 1–1.5 cm diam., with its upper part covered with verrucose to pulverulent, yellow-brown (5C2–6) volval remnants, with olivaceous tinge. Annulus subapical, fragile and fugacious.

Basidia 35–40 × 10–14 μm, clavate, 4-spored. Basidiospores [100/7/7] 8.5–11.0 × (6.5–) 7.0–8.5 μm, Q = 1.19–1.4 (–1.47), Qm = 1.3 ± 0.07, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Volval remnants on pileus composed of vertically to irregularly, loosely arranged filamentous hyphae and inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central and southwestern China (Yang 2015; this study), and Thailand (Sanmee et al. 2003, 2008).

Specimens examined: CHINA. HUNAN PROVINCE: Changsha, in a subtropical forest, date and collector unknown (HKAS 83217). YUNNAN PROVINCE: Baoshan, Longyang, in a subtropical mixed forest with Pinus yunnanensis, P. armandii and Fagaceae, altitude 1760 m, 27 July 2009, Li-Ping Tang 895 (HKAS 56852); Lushui County, altitude 850 m, 7 August 2010, Qing Cai 293 (HKAS 67855); Mengla County, in a forest with Fagaceae and Pinaceae, altitude 1200 m, 9 July 2014, Gang Wu 1299 (HKAS 89036); Nanjian County, Lingbaoshan National Forest Park, in a forest of Pinus, altitude 2243 m, 4 August 2014, Qing Cai 1213 (HKAS 83680); same location, in a forest of Pinus kesiya var. langbianensis, altitude 2243 m, 4 August 2014, Qing Cai 1214 (HKAS 83681). THAILAND. CHIANG MAI PROVINCE: Doi Suthep-Pui National Park, Chang-Kian, in a forest with Fagaceae, 11 June 2002, R. Sanmee, P. Lumyong, S. Lumyong, R. Kodsueb and W Chittrong s.n (ISOTYPE, HKAS 41153).

Commentary: Amanita siamensis was described from Thailand by Sanmee et al. (2003), and subsequently reported from China (Yang 2015). This species is characterized by its olivaceous tinged, yellow-brown basidioma densely covered with pulverulent, concolorous volval remnants, and broadly ellipsoid to ellipsoid basidiospores. As well as the Chinese collections, we also examined the isotype of A. siamensis. The basidiospores from the isotype are [20/1/1] (8.5–) 9.0–12.5 (–14.0) × (5.5–) 6.0–7.0 μm, Q = (1.38) 1.42–1.9 (–2.12), Qm = 1.67 ± 0.18, ellipsoid to elongate. Phylogenetically, A. siamensis was closely related to A. rufoferruginea Sanmee et al. (Fig. 1b). For comparisons between the two species see the commentary on A. siamensis in Yang (2015).

19. Amanita sinensis Zhu L. Yang var. sinensis, Bibl. Mycol. 170: 23 (1997).

Basidioma medium-sized to large. Pileus 7–12 cm in diam., grayish (1B1), gray (1C1) to dark gray (1D1); volval remnants on pileus verrucose, conical to floccose, gray (1C1), dark gray (1D1) to gray-brown (4C1–3); margin striate (0.1–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 10–15 cm long × 1–2.5 cm diam., dirty white (2B1) to grayish (1B1), covered with floccose, grayish (1B1), gray (1C1) to dark gray (1D1) squamules; context white (1A1), unchanging; basal bulb clavate to fusiform, 2–3.5 cm diam., lacking distinct limit between stipe and basal bulb, with its upper part covered with pulverulent to floccose, grayish (1B1), gray (1C1) to dark gray (1D1) volval remnants. Annulus apical to subapical, fragile and fugacious.

Basidia 40–65 × 11–16 μm, clavate, 4-spored. Basidiospores [200/12/11] (8.0–) 9.5–12.5 (–13.5) × 7.0–8.5 (–9.5) μm, Q = (1.14–) 1.25–1.53 (–1.63), Qm = 1.38 ± 0.09, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae scarce; inflated cells very abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, southern and southwestern China (Yang 1997, 2005, 2015; this study), Nepal (Oda et al. 2000), Japan (Oda et al. 2000; Imazeki et al. 2011), Republic of Korea (Kim et al. 2013b) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 955 m, 23 July 2017, Yan-Jia Hao 1546 (HKAS 100623). FUJIAN PROVINCE: Zhangping, in a subtropical forest, altitude 360 m, 4 August 2013, Nian-Kai Zeng 1383 (HKAS 100492). GUANGDONG PROVINCE: Zhaoqing, Dinghushan, in a subtropical forest, altitude ca. 300 m, 11 April 1981, Tai-Hui Li s.n. (GDGM 4345). GUANGXI PROVINCE: Binyang County, altitude ca. 125 m, 29 April 1970, Yu-Chen Zong et al. 34 (HMAS 35531). GUIZHOU PROVINCE: Tongren, Fanjingshan, altitude ca. 1000 m, August 1983, Xing-Liang Wu s.n. (HKAS 14456). HUNAN PROVINCE: Shimen County, Hupingshan, in a subtropical forest, altitude 230 m, date unknown, Ping Zhang 999 (HKAS 100491). SICHUAN PROVINCE: Pujiang County, Datang, in a pine forest, altitude 650 m, 25 June 1985, Ming-Sheng Yuan 1010 (HKAS 15842). YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a subtropical forest dominated by Pinus kesiya var. langbianensis, altitude 2000 m, 5 August 2013, Zhu L. Yang 5732 (HKAS 80012); Lancang County, in a subtropical forest, altitude 1500 m, 24 August 2016, LC-LJW 100 (HKAS 97604); Xundian County, Kedu, altitude 1800 m, date unknown, Qing Cai 674 (HKAS 100493); same location, altitude 1800 m, date unknown, Qing Cai 679 (HKAS 74388).

Commentary: Amanita sinensis var. sinensis was described from China by Yang (1997), and subsequently reported from Nepal, Japan, the Republic of Korea and Thailand (Oda et al. 2000; Sanmee et al. 2008; Imazeki et al. 2011; Kim et al. 2013b). It is sold as an edible mushroom in markets in Yunnan, southwestern China, although it may contain neuropsychiatric toxins. For detailed descriptions, comparisons with similar species, line drawings and images of A. sinensis var. sinensis see Yang (1997; 2005, 2015).

20. Amanita sinensis var. subglobispora Zhu L. Yang, T. H. Li & X. L. Wu, Fungal Divers. 6: 154 (2001).

Basidiospores [85/4/3] (7.5–) 8.5–10.5 (–11.0) × (6.5–) 7.5–9.0 (–10.0) μm, Q = (1.05–) 1.06–1.21 (–1.26), Qm = 1.13 ± 0.05, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China (Yang et al. 2001; Yang 2005).

Specimens examined: CHINA. HAINAN PROVINCE: Ledong County, Jianfengling, in a tropical forest, altitude 800 m, 22 April 1960, Ji-Hou Yu and Rong Liu 1103 (HMAS 30834); same location, in a tropical forest, altitude ca. 850 m, 2 September 1999, Xing-Liang Wu 17 (HKAS 34107); same location, in a tropical forest, altitude 900 m, 18 August 1999, Ming-Sheng Yuan 4329 (HOLOTYPE, HKAS 34568).

Commentary: Amanita sinensis var. subglobispora was described from Hainan Province, China (Yang et al. 2001). Because no sequences were successfully generated from the materials cited above, in this work we follow the concept of A. sinensis var. subglobispora in Yang et al. (2001) and Yang (2005). More collections from the type locality are needed for understanding the phylogeny and taxonomy of this species. For detailed descriptions, comparisons with similar species, and line drawings of A. sinensis var. subglobispora see Yang et al. (2001) and Yang (2005).

21. Amanita subfrostiana Zhu L. Yang, Bibl. Mycol. 170: 12 (1997).

Pileus 4–7 cm in diam., often red (7A8) to orange-red (6A7–8) at center, but orange (6A3–6) to yellow (4A3–5) towards margin; volval remnants on pileus pulverulent, floccose to felted, yellowish (3A5–8), yellow (4A3–5) to orange-red (6A7–8); margin striate (0.3–0.5 R), non-striate. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 6–10 cm long × 1–1.5 cm diam., cream (1A2) to yellowish (2A2–3); Stipe base clavate to subglobose to ovoid, 1–1.5 cm diam., with its upper part covered with collar-like, yellowish (2A2–3) volval remnants. Annulus submedian, persistent.

Basidia 38–55 × 10.5–12 μm, clavate, 4-spored. Basidiospores [240/8/8] (8.0–) 8.5–10.5 (–12.0) × (7.0–) 8.0–10.0 (–10.5) μm, Q = 1.0–1.13 (–1.22), Qm = 1.06 ± 0.04, globose to subglobose, inamyloid. Volval remnants on pileus composed of irregularly arranged inflated cells and filamentous hyphae. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study) and Nepal (Yang 2005, 2015; this study).

Specimens examined: CHINA. SICHUAN PROVINCE: Xichang, Daqingxiang, bought from roadside markets, altitude 2500 m, 27 July 2012, Yan-Jia Hao 650 (HKAS 76308). TIBET AUTONOMOUS REGION: Linzhi, altitude 3100 m, 20 August 1983, Yong-Ge Su s.n. (HKAS 16416). YUNNAN PROVINCE: Binchuan County, Jizushan, altitude ca. 2300 m, 1 September 1992, Yang Xiang 4 (HOLOTYPE, HKAS 32513); same location, in a subtropical forest with Pinaceae and Fagaceae, altitude 2360 m, 9 August 2014, Yan-Jia Hao 1380 (HKAS 83184); Nanhua County, in a mixed forest, altitude 2200 m, 3 August 2009, Qing Cai 83 (HKAS 58750); same location, in a mixed forest, altitude 2200 m, 3 August 2009, Li-Ping Tang 1085 (HKAS 57042); Jianchuan County, in a forest dominated by Fagaceae, altitude 2500 m, 30 August 2009, Qing Cai 182 (HKAS 58847). NEPAL. DHUNCHE–SINGOMPA: 18–20 September 1980, Y. Doi s.n. (TNS-F-176171).

Specimens of A. frostiana examined: USA. CONNECTICUT: Litchfield County, N. of Washington Depot, Bee Brook, altitude unknown, 25 July 1992, Jackie Agnew s.n. [Tulloss 7-25-92-E] (RET 064-5). MAINE: Cumberland County, Cumberland Center, altitude unknown, 20–25 July 1984, S. S. Ristich s.n. [Tulloss 7-20/25-84-SSR] (RET 238-1); same location, altitude unknown, 18 July 1984, S. S. Ristich s.n. [Tulloss 7-28-84-SSR-B] (RET 238-3). MASSACHUSETTS: locality unknown, altitude unknown, 16 August 1986, R. Youst s.n. [Tulloss 8-16-86-A] (RET 138-6); Essex County, Boxford State Forest, altitude unknown, 6 September 1990, Peter Fechko, Rob Moir and R. E. Tulloss s.n. [Tulloss 9-6-90-C] (RET 009-3). NEW HAMPSHIRE: Hillsborough County, Annett, altitude unknown, 18 August 1989, Joe Kuczinski s.n. [Tulloss 8-18-89-A] (RET 246-9). NEW YORK: Otsego County, Milford State Forest, altitude unknown, 16 August 1985, R. Harlow s.n. [Tulloss 8-16-85-B] (RET 101-2).

Commentary: Amanita subfrostiana was described from China by Yang (1997), and is also found in Nepal (Yang 2005, 2015; this study). Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. subfrostiana is closely related to A. frostiana (Peck) Sacc. Indeed, they are also similar in macroscopic appearance.

We have studied collections of A. frostiana from USA and our data indicate it differs from A. subfrostiana by its yellow-orange to orange-yellow pileus with relatively shorter striations at the pileus margin, a white stipe and volval remnants on the pileus composed of vertically arranged elements. For detailed descriptions, comparisons with similar species, line drawings and images of A. subfrostiana see Yang (1997, 2005, 2015).

22. Amanita subglobosa Zhu L. Yang, Bibl. Mycol. 170: 18 (1997).

Pileus 4–10 cm in diam., brownish (4B2–5), brown (4C3–5) to dark brown (4D3–5); volval remnants on pileus pyramidal to verrucose, white (1A1) to yellowish (2A2–4), removable; margin striate (0.1–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2), lamellulae truncate. Stipe 5–15 cm long × 0.5–2 cm diam., white (1A1) to dirty white (2B1); context white (1A1), unchanging; basal bulb subglobose, 1.5–3.5 cm diam., with its upper part covered with conical to pulverulent, yellowish (2A2–4) to brownish (4B2–5) volval remnants, often forming a collar between limit of stipe and basal bulb. Annulus subapical to submedian, white (1A1), persistent.

Basidia 44–65 × 10–13 μm, clavate, 4-spored. Basidiospores [557/27/15] (7.5–) 8.5–12.0 (–15.0) × (6.0–) 7.0–9.5 (–12.5) μm, Q = (1.05–) 1.15–1.43 (–1.69), Qm = 1.27 ± 0.08, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae fairly abundant; inflated cells abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, northeastern and southwestern China (Yang 1997, 2005, 2015; this study), India (Semwal et al. 2007), Republic of Korea (Kim et al. 2013b) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. HENAN PROVINCE: Neixiang County, altitude ca. 180 m, 1 August 2010, Xiang-Hua Wang 2555 (HKAS 100494). SICHUAN PROVINCE: Xichang, Luojishan, altitude 2000 m, 30 September 1983, Ming-Sheng Yuan 247 (HOLOTYPE, HKAS 12009). LIAONING PROVINCE: Huanren County, in a forest of Fagaceae, altitude 397 m, 26 August 2015, Qing Cai 1431 (HKAS 92050). YUNNAN PROVINCE: Baoshan, Longyang, in a forest of Pinus yunnanensis, altitude 1680 m, 11 August 2010, Qing Cai 350 (HKAS 67914); Changning County, in a forest of Pinus yunnanensis, altitude 1784 m, 24 July 2009, Li-Ping Tang 917 (HKAS 56874); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 1700 m, 25 July 2009, Li-Ping Tang 936 (HKAS 56893); Gongshan County, Bingzhongluo, in a forest with Fagaceae, altitude 2000 m, 2 August 2010, Qing Cai 250 (HKAS 67812); Jianchuan County, in a forest dominated by Pinus yunnanensis, altitude 2260 m, 29 August 2009, Qing Cai 172 (HKAS 58837); Lanping County, in a forest dominated by Pinus yunnanensis, altitude 2250 m, 15 August 2011, Gang Wu 703 (HKAS 75017); same location, altitude 2300 m, 15 August 2010, Bang Feng 858 (HKAS 68639); Longyang, in a forest dominated by Pinus yunnanensis, altitude 1243 m, 11 August 2010, Li-Ping Tang 1243 (HKAS 63063); Ninger County, in a mixed forest with Fagaceae and Pinaceae, altitude 1400 m, 1 August 2008, Li-Ping Tang 556 (HKAS 54787); Puer, in a forest with Fagaceae, altitude 1326 m, 11 July 2014, Li-Hong Han 447 (HKAS 84743); Yulong Naxi Autonomous County, in a forest of Pinus yunnanensis, altitude 2600 m, 20 August 2010, Xue-Tai Zhu 257 (HKAS 68433); Yongping County, in a mixed forest with Fagaceae and Pinaceae, altitude 2100 m, 31 July 2009, Qing Cai 69 (HKAS 58834).

Commentary: Amanita subglobosa was described from China by Yang (1997). It was subsequently reported from India, the Republic of Korea and Thailand (Semwal et al. 2007; Sanmee et al. 2008; Kim et al. 2013b).

In China, A. subglobosa has been previously misidentified as A. pantherina. They are not only similar in macroscopic appearance, but also are related to each other in our multi-locus phylogenetic analysis (Fig. 1b). However, they represent two distinct species and the differences between them were discussed in Yang (1997, 2005, 2015). For detailed descriptions, line drawings and images of A. subglobosa see Yang (1997, 2005, 2015).

23. Amanita subparvipantherina Zhu L. Yang, Qing Cai & Yang-Yang Cui, Fungal Divers. 15 (1): 194 (2015).

Pileus 5–7 cm in diam., yellowish brown (4A4–5, 4B4–7) to brown (4C5–8) or dark brown (4D6–8, 4E6–8), becoming yellowish (4A2–4) to brownish (4B3–4, 4C3–4) towards margin; volval remnants on pileus conical, subconical to granular, dirty white (2B1) to grayish (3B1–2) to brownish (3A2–4, 3B2–3); margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae truncate. Stipe 11–16.8 cm long × 0.8–2 cm diam., white (1A1); context white (1A1), unchanging; basal bulb subglobose, 2–3 cm diam., with its upper part covered with subconical to granular, grayish (3B1–2) to brownish (3A2–4, 3B2–3) volval remnants, often forming a short limbate collar on stipe base. Annulus subapical.

Basidia 30–40 × 9–13 μm, 4-spored. Basidiospores [140/11/11] (8.0–) 9.0–11.5 (–13) × (5.5–) 6.5–8.0 (–9.5) μm, Q = (1.20–) 1.28–1.5 (–1.69), Qm = 1.38 ± 0.11, broadly ellipsoid to ellipsoid, rarely elongate, inamyloid, colorless, hyaline, thin-walled, smooth; apiculus small. Volval remnants on pileus composed of somewhat irregularly arranged elements: filamentous hyphae very abundant; inflated cells fairly abundant to abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Ariyawansa et al. 2015; this study) and India (Mehmood et al. 2018).

Specimens examined: CHINA. YUNNAN PROVINCE: Binchuan County, Jizushan, in a forest dominated by Pinus yunnanensis, altitude 2000–2100 m, 11 August 2011, Qing Cai 657 (HKAS 70252); same location, in a mixed forest with Fagaceae and Pinaceae, altitude ca. 2000 m, 9 August 2014, Qing Cai 1288 (HKAS 83757); Dali, Jizushan, in a forest with Fagaceae, altitude 2300 m, 4 August 2013, Zhu L. Yang 5728 (HKAS 80008); Jingdong County, Ailaoshan, in a forest with Fagaceae, altitude 2500 m, 15 July 2008, Li-Ping Tang 333 (HKAS 54564); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 2580 m, 6 August 2014, Qing Cai 1240 (HKAS 83709); Lijiang, Lijiang Alpine Botanic Garden, altitude 2650 m, 9 August 2014, Xiao-Bin Liu 683 (HKAS 87232); Longyang, in a mixed forest with Fagaceae and Pinaceae, altitude 2400 m, 12 August 2011, Yan-Jiao Hao 485 (HKAS 71594); Lushui County, in a forest of Pinus yunnanensis, altitude 2000 m, 7 August 2010, Qing Cai 291 (HKAS 67853); Tengchong County, in a mixed forest, altitude 1800 m, 20 July 2009, Li-Ping Tang 860 (HKAS 56817); Yongping County, in a mixed forest, altitude 1991 m, 30 July 2009, Li-Ping Tang 1029 (HOLOTYPE, HKAS 56986); Yulong Naxi Autonomous County, Shitouxiang, in a forest of Picea, altitude 3000 m, 5 September 2009, Qing Cai 226 (HKAS 58891).

Commentary: Amanita subparvipantherina was described from China by Ariyawansa et al. (2015). It was subsequently reported from India (Mehmood et al. 2018). Species to which it is closely related are as yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. subparvipantherina see Ariyawansa et al. (2015).

24. Amanita sychnopyramis Corner & Bas f. sychnopyramis, Persoonia 2(3): 291 (1962).

Pileus 3–8 cm in diam., brownish (2B34), gray-brown (3C2–5) to dark brown (4C3–7); volval remnants on pileus pyramidal, white (1A1), dirty white (2B1) to grayish (4B1); margin striate (0.3–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1), lamellulae truncate. Stipe 5–11 cm long × 0.7–1.5 cm diam., white (1A1) to cream (1A2); context white (1A1), unchanging; basal bulb subglobose to fusiform, 1.5–2 cm diam., with its upper part covered with verrucose, conical to pulverulent, yellowish (4A3) to grayish (4B1) volval remnants. Annulus absent.

Basidia 30–42 × 8–12 μm, clavate, 4-spored. Basidiospores [20/1/1] (6.0–) 6.5–8.5 × (5.5–) 6.0–8.0 (–8.5) μm, Q = 1.0–1.11, Qm = 1.06 ± 0.03, globose to subglobose, inamyloid. Volval remnants on pileus composed of vertically arranged elements: filamentous hyphae scarce to fairly abundant; inflated cells very abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2015; this study), Malaysia (Lee 2017) and Singapore (Corner and Bas 1962).

Specimens examined: CHINA. YUNNAN PROVINCE: Malipo, Dongguacun, in a subtropical mixed forest with Fagaceae and Cunninghamia, altitude 1150 m, 5 August 2014, Li-Hong Han 527 (HKAS 83454).

Commentary: Amanita sychnopyramis f. sychnopyramis was described from Singapore by Corner and Bas (1962). It was subsequently reported from China and Malaysia (Yang 2015; Lee 2017).

Collections of A. sychnopyramis f. sychnopyramis and A. sychnopyramis f. subannulata split into two subclades within a monophyletic clade based on PTP analysis of our multi-locus phylogenetic data (Fig. 2). However, when more collections were included, the two subclades were not supported (Figs. 1b, S6), and collections of A. sychnopyramis and its varieties clustered within each other as a single monophyletic clade. In addition, the morphological discrepancy between them is limited to the presence of a median annulus in A. sychnopyramis f. subannulata (Hongo 1971; Imazeki and Hongo 1987; Yang 2005, 2015). Consequently, we maintain the current taxonomic ranks for A. sychnopyramis and its varieties in this study.

In our multi-locus phylogenetic analysis (Fig. 1b), A. sychnopyramis f. sychnopyramis is related to A. elata. However, A. elata differs morphologically in that it has a relatively paler pileus (yellowish-brown) covered with floccose to pulverulent volval remnants and relatively larger basidiospores (7.5–9.5 × 7–9 μm) (Corner and Bas 1962; Yang 2015). For detailed descriptions, comparisons with similar species and images of A. sychnopyramis f. sychnopyramis see Yang (2015).

25. Amanita sychnopyramis f. subannulata Hongo, Mem. Fac. Liberal Arts Shiga Univ. Nat. Sci. 21: 63 (1971).

Synonym: Amanita gleocystidiosa Boonprat. & Parnmen, Fungal Diversity 78: 136 (2016), syn. nov.

Annulus present and median, white (1A1) to cream (1A2); microscopically, dominantly composed of radially arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells rare, but becoming abundant at annular edge.

Habitat: Solitary to scattered in broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, eastern, southern and southwestern China (Yang 1997, 2005, 2015; this study) and Japan (Hongo 1971; Oda et al. 1999; Imazeki et al. 2011).

Specimens examined: CHINA. HUNAN PROVINCE: Yongzhou, Yangmingshan, in a mixed forest with Fagaceae and Pinaceae, altitude 539 m, 6 July 2012, Qing Cai 703 (HKAS 75485). FUJIAN PROVINCE: Xiamen, in a subtropical forest, altitude 600 m, 10 September 2011, Xiao-Fei Shi 938 (HKAS 101427). JIANGXI PROVINCE: Jiujiang, Lushan Scenic Spot, in a mixed forest with Fagaceae and Pinaceae, altitude 950–994 m, 21 July 2017, Gang Wu 2184 (HKAS 101438). YUNNAN PROVINCE: Xichou County, in a subtropical forest with broad-leaved trees, altitude 1550 m, 16 June 1992, Zhu L. Yang 1717 (HKAS 26141); same location, in a subtropical forest, altitude 1550 m, 16 June 1992, Zhu L. Yang 1743 (HKAS 26144). ZHEJIANG PROVINCE: Linan, Dongtianmushan, in a mixed forest with Fagaceae and Pinaceae, altitude 640–870 m, 15 July 2017, Gang Wu 2125 (HKAS 101437). JAPAN. SHIGA: Ōtsu, in a forest with Castanopsis, altitude unknown, 15 July 1970, Hongo Herb. 4166 (TNS-F-237283).

Commentary: Amanita sychnopyramis f. subannulata was described from Japan by Hongo (1971), and subsequently reported from China (Yang 1997, 2005, 2015).

In our phylogenetic analyses (Figs. 1b, S6), collections of A. sychnopyramis and its varieties clustered within each other as a monophyletic clade, and the morphological discrepancy between them is limited to the presence of a median annulus in A. sychnopyramis f. subannulata (Hongo 1971; Imazeki and Hongo 1987; Yang 2005, 2015).

In our multi-locus phylogenetic analysis (Fig. 1b), A. sychnopyramis f. subannulata is related to A. elata. However, A. elata has a relatively paler pileus (yellowish-brown) covered with floccose to pulverulent volval remnants, no annulus and relatively larger basidiospores (7.5–9.5 × 7–9 μm) (Corner and Bas 1962; Yang 2015).

In our ITS phylogenetic analysis (Fig. S6), A. gleocystidiosa clustered within the monophyletic clade of A. sychnopyramis and because of this, coupled with the presence of the annulus mentioned in Li et al. (2016), led us to treat it as a synonym of A. sychnopyramis f. subannulata. The presence of cystidia in A. gleocystidiosa (Li et al. 2016) could be a misinterpretation.

For detailed descriptions, comparisons with similar species, line drawings and images of A. sychnopyramis f. subannulata see Yang (1997, 2005, 2015).

Amanita sect. Amarrendiae (Bougher & Lebel) Zhu L. Yang, Yang-Yang Cui, Qing Cai & Li-Ping Tang, comb. & stat. nov.

MycoBank: MB 826869

Basionym: Amarrendia Bougher & T. Lebel, Aust. Syst. Bot. 15(4): 514 (2002).

Type: Amanita oleosa (Bougher & T. Lebel) Justo, Mycologia 102(3): 682 (2010).

Diagnosis: Basidiospores inamyloid; clamps present. When basidioma agaricoid, pileus covered with volval remnants; lamellulae truncate; stipe base mostly non- to slightly inflated. When basidioma sequestrate, stipitate or astipitate. Distributed in Southern Hemisphere and associated with Nothofagus and Eucalyptus.

Description: Basidioma agaricoid or sequestrate (stipitate or astipitate). When basidioma agaricoid, pileus applanate with a more or less striate margin; volval remnants on pileus present; trama white to yellow; lamellae free, white to yellow; lamellulae truncate; stipe subcylindric, stipe base non- or slightly inflated, occasionally obviously inflated; volval remnants on stipe base present; annulus mostly median and small; basidiospores inamyloid, mostly thin-walled and smooth, occasionally thick-walled and ornamented; clamps present in all parts of basidioma. When basidioma sequestrate, gleba loculate, non- or exposed, locules empty or filled with liquid; stipitate or astipitate, stipe apex mostly not or occasionally extending to the top of gleba; basidiospores statismosporic, inamyloid, mostly thin-walled and smooth, occasionally thick-walled and ornamented; basidia mostly 4-spored, or rarely 2-spored; sterigmata stout and straight; clamps present in all parts of basidioma. Currently only known in Southern Hemisphere in forests with Nothofagus, Eucalyptus, etc.

Commentary: To date, there is evidence that eight sequestrate species are members of Amanita, namely A. arenaria (O. K. Mill. & E. Horak) Justo, A. torrendii Justo, A. grandis (Bougher) Justo, A. inculta (Bougher) Justo, A. pseudoinculta Justo, A. oleosa (Bougher & T. Lebel) Justo, A. grandispora (G. W. Beaton, Pegler & T. W. K. Young) Justo, and A. nouhrae Truong, Kuhar & M. E. Sm. (Miller 1992; Malençon 1995; Bougher 1999; Bougher and Lebel 2002; Tulloss 2009; Justo et al. 2010; Truong et al. 2017). Of these sequestrate species, six (A. grandis, A. inculta, A. pseudoinculta, A. oleosa, A. grandispora and A. nouhrae) clustered with four agaricoid species (A. morenoi Raithelh., A. muriniflammea Tulloss, A. M. Young & A. E. Wood, A. umbrinella E.-J. Gilbert & Cleland and A. merxmuelleri Bresinsky & Garrido) as a distinct clade with high support (Justo et al. 2010; Truong et al. 2017; this study). Furthermore, species in this clade can be characterized by their agaricoid or sequestrate basidioma, thick-walled and ornamented, or thin-walled and smooth basidiospores, the presence of clamps and distributions through the Southern Hemisphere in forests with Nothofagus and Eucalyptus. Therefore, we treated this clade as a novel section. Bougher and Lebel (2002) proposed the genus Amarrendia typified by Amarrendia oleosa. Then Justo et al. (2010) treated Amarrendia as a synonym of Amanita located in “Amarrendia clade” without section assignment by molecular data. In our study, we named this section as Amanita sect. Amarrendiae based on the basionym of Amarrendia. Based on our phylogenetic analysis (Fig. 1a, b), A. sect. Amarrendiae was sister to A. sect. Amanita.

Although no species from A. sect. Amarrendiae were found in China, we have studied collections of A. pseudoinculta and A. umbrinella from Australia and provided morphological descriptions below, together with DNA sequences submitted to NCBI.

Amanita pseudoinculta Justo, Mycologia 102(3): 682 (2010).

Figure 7.
Fig. 7

Basidiospores of Amanita pseudoinculta (PERTH 08105731). Bars = 10 μm

Basidioma sequestrate, stipitate. Gleba loculate. Basidiospores (Fig. 7) [30/2/2] (11.0–) 13.0–16.0 × 7.5–9.5 μm, Q = (1.47–) 1.53–1.82, Qm = 1.67 ± 0.1, ellipsoid to elongate, statismosporic, inamyloid, mostly thin to slightly thick-walled (less than 0.5 μm thick). Volval remnants on pileus composed of very abundant filamentous hyphae 2–5 μm in diam., mixed with fairly abundant to abundant inflated cells, 40–80 × 30–60 μm, globose to broadly ellipsoid. Stipe trama dominantly composed of terminal inflated cells, clavate to narrowly clavate, 200–300 × 20–40 μm, mixed with fairly abundant filamentous hyphae 1.5–10 μm in diam.; vascular hyphae present but not abundant, 2–10 μm in diam. Clamps present in all parts of basidioma.

Habitat: Under trees of Eucalyptus and Acacia.

Distribution: Known from Australia.

Specimens examined: WESTERN AUSTRALIA. MOUNT GIBSON STATION: 29°36′8.400″S, 117°11′59.800″E, 17 September 2009, M. D. Barrett Bou 00579 (PERTH 08474028). RED GUM PASS: Stirling Range National Park, under Eucalyptus and Acacia, 34°24′25.400″S, 117°44′44.600″E, 16 July 2007, K. Syme 2051 (PERTH 08105731).

Commentary: Amanita pseudoinculta was described by Justo et al. (2010) as a member of the “Amarrendia clade” of Amanita. In our phylogenetic analysis (Fig. 1b), A. pseudoinculta is closely related to A. inculta in A. sect. Amarrendiae. However, A. inculta differs from A. pseudoinculta in that its gleba often fragments as the stipe elongates, and it has ellipsoid to subovoid basidiospores (9.0–14.0 × 6.0–7.0 μm, Q = 1.84) (Bougher 1999; Justo et al. 2010).

Five other sequestrate species in the Amanitaceae, A. torrendii, A. grandis, A. oleosa, A. grandispora and A. nouhrae, are also comparable to A. pseudoinculta. However, A. torrendii, a species in A. sect. Caesareae, differs from A. pseudoinculta by its narrower basidiospores (12.6–16.0 × 5.0–7.0 μm, Q = 2.37) and distribution in southern Europe and northern Africa in association with Pinus, Larix and Quercus (Malençon 1955; Miller and Horak 1992; Bougher 1999; Justo et al. 2010). Amanita grandis has a relatively larger pileus (up to 4 cm in diam.) expanding outwards from stipe exposing much of the lower gleba, and a cylindrical stipe with its apex extending through the entire gleba to the top of the pileus (Bougher 1999; Justo et al. 2010). Amanita oleosa has an astipitate basidioma, locules in the gleba filled with liquid, and relatively broader, broadly ellipsoid to subovoid basidiospores (12.5–15.0 × 9.0–11.0 μm, Q = 1.39) (Bougher and Lebel 2002; Justo et al. 2010). Amanita grandispora has an astipitate basidioma and broader, broadly ellipsoid to subovoid basidiospores (13.0–17.0 × 9.5–11.5 μm, Q = 1.43) (Bougher and Lebel 2002; Justo et al. 2010). Amanita nouhrae has distinctly ornamented, subglobose to deformed basidiospores (14.0–20.0 × 12.5–17.5 μm, Q = 1.05–1.25), is distributed in northern Patagonia and is associated with Nothofagus antarctica (Truong et al. 2017).

Amanita umbrinella E.-J. Gilbert & Cleland, in Bresadola, Iconogr. Mycol., Suppl. I 27: 273 (1941).

Pileus dark gray (4E1–5), with its surface covered with grayish (1C1–3), appressed, patchy volval remnants. Stipe annulate; upper surface annulus striate. Basidia 48–72 × 11–13.5 μm, clavate, 4-spored; sterigmata 4–5 μm long; basal septa clamped. Basidiospores [20/1/1] (9.5–) 10.5–12.5 (–13.0) × (7.5–) 8.5–10.0 (–10.5) μm, Q = (1.1–) 1.15–1.33, Qm = 1.24 ± 0.06, broadly ellipsoid, inamyloid. Clamps present in all parts of basidioma.

Habitat: On sandy ground under planted eucalypts with she oak and acacias.

Distribution: Australia.

Specimen examined: WESTERN AUSTRALIA. PERCH: Kings Park, about 200 m E. of Saw Avenue, 31°57′38.000″S, 115°49′39.300″E, 28 June 2012, N. C. Bougher 1029 (PERTH 08478090).

Commentary: Amanita umbrinella was originally described as a nomen nudum in Amanitaria by Gilbert (1940), and later formally described as a new species of Amanita without section assignment (Gilbert 1941a). In the study of Truong et al. (2017) and in our phylogenetic analysis (Fig. 1b), four agaricoid species in A. sect. Amarrendiae, including A. morenoi, A. muriniflammea, A. umbrinella and A. merxmuelleri, clustered as separated lineages and represented different species. However, their morphological characters are still insufficiently known due to limited material.

Amanita sect. Caesareae Singer ex Singer, Acta Inst. Bot. Acad. Sci. USSR, Ser. II, Plant. Crypt. 6: 398 (1951).

Synonym: Torrendia Bres., Atti Imp. Regia Accad. Rovereto, ser. 3 8: 132 (1902).

Lectotype: Amanita caesarea (Scop.) Pers., Syn. meth. fung. (Göttingen) 2: 252 (1801).

Diagnosis: For agaricoid species, pileus margin striate; lamellulae truncate; stipe base bulbless and covered with a saccate volva; annulus present; basidiospores inamyloid and clamps present. For the unique sequestrate and stipitate species, A. torrendii, basidiospores inamyloid and clamps present.

Commentary: This study reports 20 species of Amanita sect. Caesareae from China, including six novel species and 14 known species. Of the known taxa, A. hemibapha var. ochracea was elevated to species rank as A. ochracea. To date, no sequestrate taxa of this section have been reported from China. Previous studies and our multi-locus phylogenetic data indicate that the genus Torrendia is a synonym of A. sect. Caesareae (Fig. 1b) (Bas 1975; Tulloss 2009; Justo et al. 2010).

Key to the species of Amanita sect. Caesareae from China

  1. 1.

    Basidiospores subglobose to broadly ellipsoid, 9.0–13.0 × 8.5–11.0 μm, Q = 1.0–1.33……….2

     
  2. 1.

    Basidiospores broadly ellipsoid, ellipsoid to elongate, 8.0–13.0 × 5.5–10.0 μm, Q = 1.2–1.6……….7

     
  3. 2.

    Pileus white, often yellowish to cream at center……….3

     
  4. 2.

    Pileus gray, grayish to gray-brown or brown to yellow-brown……….4

     
  5. 3.

    Striations on pileal margin relatively longer (0.3–0.4 R); annulus persistent and becoming yellow when mature; basidiospores globose to subglobose, 9.0–11.0 × 9.0–11.0 μm, Q = 1.0–1.13, Qm = 1.06 ± 0.03………A. alboumbelliformis

     
  6. 3.

    Striations on pileus margin relatively shorter (0.2–0.3 R); annulus fragile and white; basidiospores subglobose to broadly ellipsoid, 9.5–12.5 × 8.5–11.5 μm, Q = 1.07–1.28, Qm = 1.17 ± 0.06………A. chepangiana

     
  7. 4.

    Basidioma relatively smaller with pileus 6.5–15 cm in diam.; outer surface of volval remnants on stipe base not cracking into brownish patches……….5

     
  8. 4.

    Basidioma relatively larger with pileus ca. 10–20 cm in diam.; outer surface of volval remnants on stipe base cracking into brownish patches………A. princeps

     
  9. 5.

    Basidioma relatively larger with pileus ca. 7–15 cm in diam.; stipe white, sometimes covered with indistinct fibrils……….6

     
  10. 5.

    Basidioma relatively smaller with pileus ca. 6.5 cm in diam.; stipe densely covered with distinct, gray, gray-brown to dark gray squamules………A. squarrosipes

     
  11. 6.

    Pileus gray, grayish to gray-brown; pileal margin with relatively shorter striations (0.1–0.2 R………A. imazekii

     
  12. 6.

    Pileus brownish, brown to yellow-brown; pileal margin with relatively longer striations (0.2–0.3 R………A. pseudoprinceps

     
  13. 7.

    Lamellar edge yellowish, yellow, red to orange-red; stipe surface covered with red, yellow, orange to brown squamules; pileus bright-colored, yellow, yellow-brown, brown, orange to red……….8

     
  14. 7.

    Lamellar edge white to pale grayish; stipe surface covered with white to gray squamules; pileus not bright-colored, gray to gray-brown……….16

     
  15. 8.

    Basidioma yellow, yellow-brown to brown, lacking red tinge……….9

     
  16. 8.

    Basidioma orange-yellow, orange to orange-red, with obviously red tinge……….12

     
  17. 9.

    Basidioma medium-sized to large; pileus ca. 6–12 cm in diam., non- or umbonate at center; basidiospores relatively smaller, ellipsoid, 8.5–10.5 × 6.0–7.5 μm; basidioma distributed in tropical to subtropical forests……….10

     
  18. 9.

    Basidioma large to very large; pileus ca. 10–25 cm in diam, umbonate at center; basidiospores relatively larger, broadly ellipsoid to ellipsoid, 9.0–12.5 × 7.0–9.0 μm; basidioma distributed in subalpine forests………A. ochracea

     
  19. 10.

    Pileus gray-brown to brown at center, yellow at margin; basidioma distributed in tropical to subtropical forests dominated by broad-leaved trees……….11

     
  20. 10.

    Pileus yellow over the entire disk; basidioma distributed in subtropical pine or mixed forests………A. kitamagotake

     
  21. 11.

    Stipe covered with yellow squamules; annulus yellow………A. fuscoflava

     
  22. 11.

    Stipe covered with orange squamules; annulus orange………A. similis

     
  23. 12.

    Pileus non-umbonate at center……….13

     
  24. 12.

    Pileus umbonate at center……….15

     
  25. 13.

    Lamellar edge often yellowish to yellow; annulus yellow to yellowish……….14

     
  26. 13.

    Lamellar edge often reddish to reddish orange; annulus red, orange-red to orange………A. rubromarginata

     
  27. 14.

    Pileus orange at center, and yellow to yellowish at margin; basidiospores relatively broader, 8.0–11.0 × 6.0–8.0 μm………A. subhemibapha

     
  28. 14.

    Pileus red at center, and orange to yellow at margin; basidiospores relatively narrower, 8.0–10.0 × 5.5–6.5 μm………A. hemibapha

     
  29. 15.

    Pileus red to dark red at center, and yellow at margin, with obvious color differences between center and margin; basidioma dominant in subtropical broad-leaved or mixed forests………A. rubroflava

     
  30. 15.

    Pileus red to orange-red, and orange-red to orange at margin, without distinct color differences between center and margin; basidioma dominant in temperate broad-leaved forests………A. caesareoides

     
  31. 16.

    Lamellae pinkish to reddish……….17

     
  32. 16.

    Lamellae white to cream……….18

     
  33. 17.

    Pileus margin with relatively shorter striations (0.2–0.4 R); basidiospores relatively narrower, 10.0–13.0 × 7.0–9.0 μm, Q = 1.31–1.5, Qm = 1.4 ± 0.07; basidioma dominant in pine, broad-leaved or mixed forests………A. incarnatifolia

     
  34. 17.

    Pileus margin with relatively longer striations (0.3–0.5 R); basidiospores relatively wider, 10.0–13.0 × 8.0–11.0 μm, Q = 1.11–1.3, Qm = 1.21 ± 0.08; basidioma dominant in pine forests………A. longistriata

     
  35. 18.

    Pileal surface with innate spots or speckles……….19

     
  36. 18.

    Pileal surface lacking innate spots or speckles………A. esculenta

     
  37. 19.

    Pileal margin with relatively longer striations (0.2–0.3 R); stipe surface covered with gray-brown squamules; volval remnants on stipe base composed of abundant, filamentous hyphae and inflated cells………A. hunanensis

     
  38. 19.

    Pileal margin with relatively shorter striations (ca. 0.1 R); stipe surface covered with white to pale gray squamules; volval remnants on stipe base composed of very abundant, filamentous hyphae and rare, inflated cells………A. yuaniana

     

26. Amanita alboumbelliformis Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 3f, g, 8.
Fig. 8

Microscopic features of Amanita alboumbelliformis. a Hymenium and subhymenium (TYPE, HKAS 83448); b basidiospores (TYPE, HKAS 83448); c longitudinal section of outer part of volval remnants on stipe base, left side indicates outer surface of volval remnants on stipe base (HKAS 100495). Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 826873

Etymology: alboumbelliformis, from albus = white, and umbelliformis = umbrella-like, referring to the white umbrella-like basidioma of this species.

Type: CHINA. YUNNAN PROVINCE: Qiubei County, in a subtropical mixed forest with Fagaceae, Pinus and Keteleeria fortune, altitude 1569 m, 10 August 2014, Li-Hong Han 593 (TYPE, HKAS 83448, as A. cf. chepangiana in Yang 2015, GenBank Acc. No.: nrLSU = MH486635, tef1-α = MH508892, rpb2 = MH486085, β-tubulin = MH485598).

Basidiomata (Fig. 3f, g) medium-sized to large. Pileus 7–10 cm diam., plano-convex to applanate, sometimes umbonate at center, white (1A1), but cream (1A2–4) at center; volval remnants absent; margin striate (0.3–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (1A2–4); lamellulae truncate, plentiful. Stipe 12–16 cm long × 1.5–2 cm diam., subcylindric or slightly tapering upward, with apex slightly expanded, white (1A1), with its surface covered with white (1A1) to cream (1A2–4) fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 5–8 cm high × 4–5 cm wide, membranous, outer surface white (1A1), inner surface white (1B1) to grayish (2B2–4). Annulus persistent, subapical, white (1A1) at young stage, but becoming yellow (1A2–3) when mature, membranous. Odor indistinct.

Lamellar trama bilateral. Mediostratum 35–50 μm wide, composed of abundant fusiform, ellipsoid to clavate inflated cells (40–110 × 10–45 μm); filamentous hyphae abundant, 2–9 μm wide; vascular hyphae rare. Lateral stratum composed of abundant, ellipsoid to clavate inflated cells (20–70 × 10–20 μm), diverging at an angle of ca. 30° to 45° to the mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 8a) 30–50 μm thick, with 2–3 layers of subglobose, fusiform, ellipsoid to irregularly arranged cells, 10–20 × 9–15 μm. Basidia (Fig. 8a) 50–75 × 14–18 μm, clavate, 4-spored; sterigmata 4–7 μm long; basal septa clamped. Basidiospores (Fig. 8b) [120/3/2] 9.0–11.0 (–13.0) × (8.0–) 9.0–11.0 (–12.0) μm, Q = 1.0–1.13 (–1.16), Qm = 1.06 ± 0.03, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose or sphaeropedunculate inflated cells (10–40 × 10–30 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 55–110 μm thick; upper layer (30–70 μm thick) gelatinized, composed of radially arranged to interwoven, thin-walled, colorless, filamentous hyphae 2–7 μm wide; lower layer (20–60 μm thick) composed of radially arranged, filamentous hyphae 2–10 μm wide, colorless to brownish; vascular hyphae rare. Interior of volval remnants on the stipe base (Fig. 8c) dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 2–9 μm wide, colorless to yellowish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to locally abundant, subglobose, subfusiform to ellipsoid, 30–90 × 15–50 μm, colorless to yellowish, thin-walled, terminal or in chains of 2–3; vascular hyphae rare. Outer (Fig. 8c) and inner surfaces of volval limb similar to the structure of interior part, but inner surface gelatinized. Stipe trama composed of longitudinally arranged, long clavate terminal cells, 50–350 × 15–60 μm; filamentous hyphae scattered to abundant, 3–10 μm wide; vascular hyphae rare. Annulus dominantly made of two parts intergrading to each other. Outer part of annulus made of radially arranged to interwoven elements: inflated cells rare to scattered, subglobose, fusiform to ellipsoid, 20–40 × 15–30 μm, pale yellowish, thin-walled; filamentous hyphae very abundant to dominant, 2–7 μm wide, pale yellowish, thin-walled; vascular hyphae rare. Inner part of annulus composed of radially arranged elements: inflated cells abundant, ellipsoid to clavate, 40–110 × 10–30 μm, pale yellowish, thin-walled; filamentous hyphae very abundant, 2–9 μm wide, pale yellowish, thin-walled; vascular hyphae rare. Clamps present in all parts of basidiomata.

Habitat: Solitary to scattered on soil in in subtropical to tropical forests associated with mixed or broad-leaved trees; basidioma occurring in summer and autumn.

Distribution: Known from southwestern and southern China.

Additional specimen examined: CHINA. FUJIAN PROVINCE: in a broad-leaved forest, 9 August 2013, Nian-Kai Zeng 1397 (HKAS 100495).

Commentary: Amanita alboumbelliformis is characterized by its white basidioma with both its pileus center and annulus cream to yellow, a long-striate pileus margin, globose to subglobose basidiospores (9–11 × 9–11 μm), the presence of clamps and an association with mixed or broad-leaved trees in subtropical forests.

Our multi-locus phylogenetic analysis (Fig. 2) suggested that A. alboumbelliformis is sister to A. aff. princeps (RET 357-5), which was contributed by Sánchez–Ramírez et al. (2015). Whether A. aff. princeps (RET 357-5) represents A. alboumbelliformis or a new taxon needs more collections and further morphological study.

Morphologically, A. alboumbelliformis is similar to A. chepangiana in their white to cream basidiomata. However, A. chepangiana has relatively shorter striations on pileal margin ca. 0.2–0.3 R, a fragile, white annulus and subglobose to broadly ellipsoid basidiospores (9.5–12.5 × 8.5–11.5 μm, Q = 1.07–1.28, Qm = 1.17 ± 0.06) (Tulloss and Bhandary 1992; Yang 2000; this study).

The European A. caesarea f. alba (Gillet) E. J. Gilbert is also comparable to A. alboumbelliformis, but the former has a thicker pileus with shorter striations on its margin, a shorter and thicker stipe, and narrower and longer basidiospores (10.0–12.0 × 7.0–8.5 μm) (Galli 2001; Neville and Poumarat 2004).

Amanita egregia D. A. Reid and A. loosei Beeli are also similar to A. alboumbelliformis. However, A. egregia differs from A. alboumbelliformis by the relatively larger basidioma with pileus that is ca. 10–20 cm in diam., and its open eucalypt forest habitat (Reid 1978, 1980). Amanita loosei has relatively shorter striations on the pileal margin, a pure white annulus and relatively longer basidiospores (Gilbert 1940; Walleyn and Verbeken 1998).

27. Amanita caesareoides Lj. N. Vassiljeva, Notul. syst. Sect. cryptog. Inst. bot. Acad. Sci. U. S. S. R. 6: 199 (1950).

Pileus 5–13 cm in diam., applanate, often umbonate at center, red (9A7–8), orange-red (7A5–8) to orange (6A6–8); volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate. Lamellae cream (1A2–4) to yellowish (3A2–6); lamellulae truncate. Stipe 8–18 cm long × 0.7–2 cm diam., yellowish (3A2–6) to yellow (4A4–7), with its surface covered with snakeskin-shaped, orange (6A6–8) squamules; basal bulb absent; volval remnants on stipe base saccate. Annulus subapical, yellow (4A4–7).

Basidia 33–45 × 8–12 μm, clavate, 4-spored. Basidiospores [110/6/6] (7.0–) 7.5–10.0 (–11.0) × (6.0–) 6.5–8.0 (–8.5) μm, Q = (1.07–) 1.14–1.35 (–1.46), Qm = 1.25 ± 0.07, broadly ellipsoid, rarely ellipsoid or subglobose, inamyloid. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in temperate broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from northeastern China (Yang 2015; this study), India (Bhatt et al. 2003, 2017), Japan (Imazeki and Hongo 1987; Endo et al. 2016), Republic of Korea (Cho et al. 2015) and Russian Far East (Vassiljeva 1950).

Specimens examined: CHINA. HEILONGJIANG PROVINCE: Fuyuan, in a temperate broad-leaved forest, altitude unknown, 5 August 2004, Bau Tolgor s. n. (HMJAU 2623). LIAONING PROVINCE: Benxi, in a temperate broad-leaved forest, altitude 432 m, 21 August 2015, Qing Cai 1397 (HKAS 92017); same location, in a forest with Fagaceae, altitude 432 m, 21 August 2015, Qing Cai 1388 (HKAS 92009). JAPAN. HOKKAIDO: Tomakomai, in a forest with Quercus and Ulmus, altitude 100 m, 12 September 2011, Zhu L. Yang 5536 (HKAS 71021); Ibarakiken, Tsukuba Botanical Garden, in a forest dominated by Fagus and Quercus, altitude 100 m, 16 September 2011, Zhu L. Yang 5573 (HKAS 71058). RUSSIA. PRIMOESKY TERRITORY: Khasansky District, Kedrovaya Pad Reserve, in an oaken forest under Quercus mongolica, altitude unknown, 22 August, E.M. Bulakh (VLAM-3904).

Specimens of A. caesarea examined: GERMANY. FRANKFURT AM MAIN: bought from a supermarket (imported to Germany from Italy), 4 October 2016, Zhu L. Yang 5968 (HKAS 96166). ITALY. TRIEST: exposition in Redipuglia, altitude unknown, 2 October 1982, C. Bas 7989 (L). AUSTRIA. BURGENLAND: Schandorf, altitude unknown, 28 July 2005, G. Koller s.n. (HKAS 56563).

Commentary: Amanita caesareoides was described from the Russian Far East by Vassiljeva (1950), and subsequently reported from China, India, Japan and the Republic of Korea (Imazeki and Hongo 1987; Bhatt et al. 2003; Cho et al. 2015; Yang 2015; Endo et al. 2016; Bhatt et al. 2017). We also studied a collection of A. caesareoides from Russia, VLAM-3904. The basidiospores of this collection were [30/1/1] (7.5–) 8.0–10.5 (–11.0) × (6.0–) 6.5–8.0 (–8.5) μm, Q = 1.13–1.33 (–1.38), Qm = 1.26 ± 0.07, mostly ellipsoid, rarely ellipsoid or subglobose. Unfortunately, we were unable to generate DNA sequences from it.

Amanita caesareoides is phylogenetically closely related and morphologically similar to the European A. caesarea (Fig. 1b). Examination of European specimens of A. caesarea has revealed that Amanita caesarea can be distinguished from A. caesareoides by its non-umbonate pileus with relatively shorter striations on its margin, a short, robust stipe and larger basidiospores (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; this study). The basidiospores of A. caesarea were [136/7/3] (9.0–) 9.5–12.0 (–14.5) × (5.5–) 6.0–7.5 (–9.0) μm, Q = (1.36–) 1.43–1.77 (–1.91), Qm = 1.59 ± 0.11, ellipsoid to elongate. For detailed descriptions, comparisons with similar species and images of A. caesareoides see Yang (2015).

28. Amanita chepangiana Tulloss & Bhandary, Mycotaxon 43: 25 (1992).

Pileus 10–14 cm in diam., white (1A1), often yellowish (3A2–4) to cream (1A2–4) at center; volval remnants on pileus absent; margin striate (0.2–0.3 R), non-appendiculate. Lamellae white (1A1), lamellulae truncate. Stipe 14–20 cm long × 1–3.5 cm diam., white (1A1); basal bulb absent; volval remnants on stipe base saccate. Annulus superior to subapical, white, fugacious.

Basidia 40–55 × 12–14.5 μm, clavate, 4-spored. Basidiospores [80/4/4] 9.5–12.5 (–14.5) × (8.0–) 8.5–11.5 (–13.5) μm, Q = 1.07–1.28 (–1.33), Qm = 1.17 ± 0.06, subglobose to broadly ellipsoid, inamyloid. Interior of volval remnants on stipe base dominantly composed of longitudinally arrange elements: filamentous hyphae abundant; inflated cells fairly abundant to abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine or broad-leaved forests (e.g. Fagaceae, Shorea robusta and Pinus); basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (this study), India (Semwal et al. 2014), Nepal (Tulloss and Bhandary 1992; Aryal and Budathokio 2012, 2013) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. YUNNAN PROVINCE: Puer, in a forest of Pinus, altitude 1370 m, 12 July 2009, Li-Ping Tang 761 (HKAS 56718). NEPAL. CHITWAN DISTRICT: Jugedi, in a Shorea robusta forest, altitude unknown, July 1986, Hemanta Ram Bhandary s.n. (PARATYPE, RET 075-7); same location, in a Shorea robusta forest, altitude unknown, 12 July 1990, Hemanta Ram Bhandary Ha p.p. (ISOTYPE, RET 259-8); same location, in a Shorea robusta forest, altitude unknown, 12 July 1990, Hemanta Ram Bhandary Hb p.p. (RET 259-9).

Commentary: Amanita chepangiana was described from Nepal by Tulloss and Bhandary (1992), and subsequently reported from India and Thailand (Sanmee et al. 2008; Semwal et al. 2014). Our phylogenetic and morphological evidence suggest the species is also found in China (Fig. 1b). HKAS 59783, which was treated as A. chepangiana in Yang (2015), represents an undescribed taxon (unpublished data) and will be described in the future when adequate collections have been made. HKAS 59783 can be distinguished from A. chepangiana by the saccate volval remnants on the stipe base, often with the outer surface cracking into grayish to brownish patches.

29. Amanita esculenta Hongo & I. Matsuda, Journ. Jap. Bot. 30 (5): 148 (1955).

Pileus 5–10 cm in diam., brown (3B2–5), brownish (1B2–4), dark gray (2A3–5) to gray-brown (4D4–6); volval remnants on pileus absent; margin striate (ca. 0.1 R), non-appendiculate. Lamellae white (1A1) to cream (1A2); lamellulae truncate. Stipe 9–12 cm long × 0.8–2 cm diam., grayish (1B1), covered with gray (2B1–3) to gray-brown (3B1–3) squamules; basal bulb absent; volval remnants on stipe base saccate. Annulus subapical, grayish (1B1–2).

Basidia 50–75 × 12–15 μm, clavate, 4-spored. Basidiospores [50/4/4] (9.5–) 10.0–12.0 × (7.0–) 7.5–9.0 μm, Q = (1.22–) 1.24–1.45 (–1.57), Qm = 1.35 ± 0.09, ellipsoid, inamyloid. Interior of volval remnants on stipe base dominantly composed of irregularly arrange elements: filamentous hyphae very abundant; inflated cells scarce to locally abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2005, 2015; this study) and Japan (Matsuda and Hongo 1955; Imazeki and Hongo 1987; Imazeki et al. 2011).

Specimens examined: CHINA. SICHUAN PROVINCE: Xichang, Daqingxiang, in a forest of Pinus yunnanensis, altitude 2000 m, 21 September 1999, Zhu L. Yang 2703 (HKAS 34169). YUNNAN PROVINCE: Lancang County, in a mixed forest with Fagaceae and Pinaceae, altitude 1201 m, 21 August 2016, LC-Hao 43 (HKAS 96880); Mengla County, in a mixed forest with Fagaceae and Pinaceae, altitude 1200 m, 9 July 2014, Gang Wu 1298 (HKAS 89035). JAPAN. HONSHU: Niigata-ko, Shunji-mura, in a pine forest, altitude unknown, 15 June 1954, Hongo Herb. 981 (TNS-F-237541).

Commentary: Amanita esculenta was described from Japan by Matsuda and Hongo (1955), and subsequently found in China (Yang 2005, 2015). In our phylogenetic analyses (Figs. 1b, S7), A. esculenta was found to be related to A. hunanensis Y. B. Peng & L. J. Liu, A. yuaniana Zhu L. Yang and A. spreta (Peck) Sacc. However, these species can be distinguished from A. esculenta through the following characters. A. hunanensis and A. yuaniana both have a pileus with distinct innate spots or speckles (Peng and Liu 1981; Yang 1994, 1997, 2005, 2015; Yang and Zhang 2002). Amanita spreta, described from the USA, differs from A. esculenta by its relatively paler colored pileus (whitish or pale brown) and narrower, elongate to cylindric basidiospores (Murrill 1913; Coker 1917; Jenkins 1978, 1986; Tulloss et al. 1995). For detailed descriptions, comparisons with similar species, line drawings and images of A. esculenta see Yang (2005, 2015).

30. Amanita fuscoflava Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 3h, 9.
Fig. 9

Microscopic features of Amanita fuscoflava (TYPE, HKAS 59800). a Hymenium and subhymenium; b basidiospores. Bars = 10 μm

MycoBank: MB 824970

Etymology: fuscoflava, from fusco- = dark brown, flavus = yellow, referring to its pileus with dark brown center and yellow margin.

Type: CHINA. HAINAN PROVINCE: Qiongzhong County, Limushan, in a tropical to subtropical forest, altitude 500 m, 5 August 2010, Nian-Kai Zeng 848 [TYPE, HKAS 59800, as Amanita sp. 5 in Yang (2015), GenBank Acc. No.: ITS = MH508372, nrLSU = MH486557, tef1-α = MH508827, rpb2 = MH486023, β-tubulin = MH485543].

Basidioma (Fig. 3h) medium-sized to large. Pileus 7–12 cm diam., convex, plano-convex to applanate, umbonate at center, dark brown (4E3–6) at center, yellow (3B3–6) to honey yellow (2A4–7) towards margin; volval remnants on pileus absent; margin striate (0.5–0.7 R), non-appendiculate; trama white (1A1) to cream (1A2), unchanging. Lamellae free, crowded, yellowish (1A3); lamellar edges cream (1A2) to yellowish (1A3); lamellulae truncate, plentiful. Stipe 8–15 cm long × 1–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, yellow (3A3–5), covered with yellow (3A3–5) to yellowish (1A3), irregularly arranged, snakeskin-shaped squamules; context white (1A1) to cream (1A2), hollow in center; basal bulb absent; volva saccate, 2–4 cm high × 2–3 cm wide, membranous, both surfaces white (1A1). Annulus subapical, yellow (3A3–5), membranous. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–40 μm wide, composed of abundant subglobose to ellipsoid inflated cells (25–60 × 15–55 μm); filamentous hyphae abundant, 4–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid, fusiform to clavate inflated cells (35–80 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–5 μm wide. Subhymenium (Fig. 9a) 30–60 μm thick, with 2–3 layers of subglobose, ovoid, fusiform, ellipsoid to irregularly arranged cells, 10–25 × 8–25 μm. Basidia (Fig. 9a) 35–48 × 10–12 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa clamped. Basidiospores (Fig. 9b) [40/2/2] 8.5–10.5 (–11.0) × (5.5–) 6.0–7.0 μm, Q = 1.31–1.64 (–1.81), Qm = 1.5 ± 0.14, ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (8–40 × 8–20 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 40–75 μm thick; upper layer (20–50 μm thick) gelatinized, composed of radially arranged to interwoven, thin-walled, colorless, filamentous hyphae 2–6 μm wide; lower layer (25–40 μm thick) composed of radially arranged, filamentous hyphae 4–9 μm wide, colorless; vascular hyphae scarce. Interior of volval remnants on stipe base dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–13 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to locally fairly abundant, subglobose, fusiform, ellipsoid to elongate, 40–90 × 20–40 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Stipe trama composed of longitudinally arranged, long clavate terminal cells, 60–400 × 15–40 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Annulus composed of radially arranged elements: inflated cells scarce, subglobose, ellipsoid, fusiform to clavate, 15–50 × 10–20 μm, colorless, thin-walled; filamentous hyphae very abundant to dominant, 1–10 μm, colorless, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in tropical to subtropical forests dominated by Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China.

Additional specimen examined: CHINA. HAINAN PROVINCE: Ledong County, Jianfengling, in a tropical to subtropical forest, altitude ca. 900 m, 27 July 2009, Xing-Liang Wu 8300 (HKAS 59742).

Commentary: Amanita fuscoflava is well characterized by its long striate pileus with a dark brown center and a yellow margin, ellipsoid basidiospores, the common presence of clamps, and its tropical distribution.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. fuscoflava is closely related to A. subhemibapha. However, the latter can be distinguished by its orange to orange-yellow umbonate pileus, relatively shorter striations on the pileal margin (0.25–0.35 R) and an orange-yellow annulus.

Amanita fuscoflava is also macroscopically similar to A. similis Boedijn, A. malayensis Tang et al. and A. aporema Boedijn. However, A. similis has a yellow stipe covered with pinkish orange squamules and a pinkish orange to grayish annulus (Boedijn 1951; Corner and Bas 1962; Lee 2017; Tang et al. 2018). Amanita malayensis has a yellow pileus with its center often yellowish orange, maize yellow or saffron yellow, shorter striations on the pileal margin (0.3–0.4 R), whitish lamellae, and slightly larger basidiospores (9.0–12.0 × 6.0–8.0 μm) (Tang et al. 2018). Amanita aporema has a non-umbonate and slightly depressed pileal center, whitish to cream lamellae, a dirty white to cream stipe, a whitish to cream annulus and globose basidiospores (9.5–11.0 × 8.5–10.5 μm) (Boedijn 1951; Lee 2017; Tang et al. 2018; this study).

Amanita chatamagotake N. Endo & A. Yamada and A. hayalyuy D. Arora & G. H. Shepard are comparable to A. fuscoflava. However, A. chatamagotake has relatively shorter striations on the pileal margin (0.2–0.5 R) (Endo et al. 2017). Furthermore, A. chatamagotake exhibits a variable pileus color. Typically, A. chatamagotake has a pileus that is grayish to olive brown, which is darker than that of A. fuscoflava (Endo et al. 2017). Amanita hayalyuy has a relatively larger basidioma with pileus ca. 8–18 cm, shorter striations on the pileal margin, and a yellow stipe decorated with orangey brown squamules (Shepard et al. 2008).

31. Amanita hemibapha (Berk. & Broome) Sacc., Syll. fung. (Abellini) 5: 13 (1887).

Basionym: Agaricus hemibaphus Berk. & Broome, Trans. Linn. Soc. London 27: 149 (1870) [1871].

Pileus 8–15 cm in diam., applanate, lacking umbo at center, red (7A6–8) to orange-red (6A6–8) at center, becoming yellow (3A6–8) towards margin; volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate. Lamellae yellowish (1A6–8) to cream (1A2–4); lamellulae truncate. Stipe 12–16 cm long × 0.5–1.5 cm diam., yellowish (1A6–8) to cream (1A2–4), covered with yellow (3A6–8) to orange-yellow (5A3–6), irregularly arranged, snakeskin-shaped squamules; basal bulb absent; volval remnants on stipe base saccate, white (1A1). Annulus subapical, yellow (3A6–8).

Basidia 35–40 × 8.5–10 μm, clavate, 4-spored. Basidiospores [85/4/3] (6.5–) 7.5–10.0 (–11.0) × (5.0–) 5.5–7.5 μm, Q = (1.2–) 1.3–1.69 (–1.72), Qm = 1.49 ± 0.1, ellipsoid, rarely elongate, inamyloid, colorless, thin-walled, smooth, apiculus small. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical forests of broad-leaved or pine trees; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study), India (Vrinda et al. 2005; Upadhyay et al. 2008), Sri-lanka (Pegler 1986) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. YUNNAN PROVINCE: Ninger County, in a forest of Pinus kesiya var. langbianensis, altitude 1400 m, 1 August 2008, Li-Ping Tang 553 (HKAS 54784). INDIA. KERALA STATE: Thiruvananthapuram District, TBGRI Campus, 30 June 2003, CK Pradeep (TBGT 5872). SRI LANKA. KANDY DISTRICT: Peradeniya, September 1868, G.H.K. Thwaites 700 [ISOTYPE of Herb. KEW, K 48990].

Commentary: Amanita hemibapha was described from Sri Lanka, and subsequently reported from China, India and Thailand (Pegler 1986; Yang 1997, 2005, 2015; Vrinda et al. 2005; Upadhyay et al. 2008; Sanmee et al. 2008). This species is characterized by its non-umbonate pileus with a red to orange-red center and a yellow and long-striate margin, a yellowish stipe covered with yellow to orange-yellow squamules, a yellow annulus, a white saccate volva attached at the very base of the stipe and ellipsoid basidiospores (Gilbert 1941a; Pegler 1986; Yang 1997, 2005, 2015; Vrinda et al. 2005; Sanmee et al. 2008). The basidiospores of the isotype of A. hemibapha were [60/2/1] (7.0–) 8.0–10.0 × (5.0–) 5.5–6.5 (–7.0) μm, Q = (1.21–) 1.36–1.64 (–1.73), Qm = 1.49 ± 0.09.

The old name A. hemibapha has hidden a species complex in Asia (Yang 2005, 2015; Upadhyay et al. 2008; Sánchez-Ramírez et al. 2015; Endo et al. 2016; Tang et al. 2018). To date, six species, A. caesareoides, A. hemibapha, A. javanica (Corner & Bas) T. Oda et al., A. rubroflava Yang-Yang Cui et al., A. rubromarginata Har. Takah. and A. subhemibapha Zhu L. Yang et al., have been recognized from this species complex in China. For detailed descriptions, comparisons with similar species and images of A. hemibapha see Yang (1997, 2005, 2015).

32. Amanita hunanensis Y. B. Peng & L. J. Liu, Acta Microbiol. Sinica 21(2): 152 (1981).

Pileus 5–12 cm in diam., gray (2B1–3) to brown (2C2–5), with innate spots or speckles; volval remnants on pileus absent, or retained as several, minute, white (1A1) patches; margin striate (0.2–0.3 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 7–15 cm long × 1–2 cm diam., white (1A1), covered with snakeskin-shaped, gray (2B1–3) to gray-brown (3C2–5) squamules; basal bulb absent; volval remnants on stipe base saccate, white (1A1) to dirty white (2B1). Annulus subapical, dirty white (2B1).

Basidia 44–65 × 10–15 μm, clavate, 4-spored; basal septa clamped. Basidiospores [70/2/2] (8.0–) 9.5–12.5 (–15.5) × 6.5–8.5 (–10.0) μm, Q = (1.22–) 1.29–1.62 (–1.71), Qm = 1.47 ± 0.1, ellipsoid, inamyloid. Interior of volval remnants on stipe base dominantly composed of very abundant inflated cells and abundant filamentous hyphae. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central China (Peng and Liu 1981; Yang 2005, 2015; this study) and India (Upadhyay et al. 2008).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 900 m, 25 July 2017, Yan-Jia Hao 1566 (HKAS 100632). HUNAN PROVINCE: Xupu County, in a forest of Pinus massoniana, altitude 400 m, 25 May 1975, Yan-Bin Peng and Lin-Han Liu 1224 (ISOTYPE, HMAS 47351).

Commentary: Amanita hunanensis was described from China by Peng and Liu (1981), and subsequently reported from India (Upadhyay et al. 2008). Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. hunanensis is closely related to A. yuaniana. However, A. yuaniana differs from A. hunanensis in its relatively shorter striations on the pileal margin (ca. 0.1 R) and scarce inflated cells in the volval remnants on the stipe base (Yang 1994, 1997, 2005, 2015).

Amanita esculenta and A. spreta are macroscopically similar to A. hunanensis. However, A. esculenta and A. spreta both have a pileus that lacks innate speckles (Murrill 1913; Coker 1917; Matsuda and Hongo 1955; Jenkins 1978, 1986; Imazeki and Hongo 1987; Tulloss et al. 1995; Yang 2005, 2015; Imazeki et al. 2011). For detailed descriptions, comparisons with similar species, line drawings and images of A. hunanensis see Yang (2005, 2015).

33. Amanita imazekii T. Oda, C. Tanaka & Tsuda, Mycologia 93(6): 1231 (2001).

Pileus 7–15 cm in diam., gray (1C1–3), grayish (1B1–3) to brown-gray (2C2–5), often darker at center, occasionally purely white (1A1); volval remnants on pileus absent; margin shortly striate (0.1–0.2 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 10–20 cm long × 1–3 cm diam., white (1A1), glabrous or covered with grayish (1B1–3) fibrils; basal bulb absent; volval remnants on stipe base saccate, white (1A1) to dirty white (2B1). Annulus subapical, white (1A1).

Basidia 48–60 (–66) × 10–17 μm, clavate, 4-spored, rarely 2-spored. Basidiospores [210/7/4] (8.5–) 9.0–12.5 (–15.0) × (8.0–) 8.5–11.5 (–13.0) μm, Q = (1.0–) 1.05–1.17 (–1.25), Qm = 1.11 ± 0.05, subglobose, sometimes broadly ellipsoid, inamyloid. Interior of volval remnants on stipe base dominantly composed of longitudinally arranged to interwoven elements: filamentous hyphae very abundant; inflated cells scarce to fairly abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn

Distribution: Known from central, northeastern and southwestern China (Yang 2005, 2015; this study), Japan (Oda et al. 2001) and Republic of Korea (Cho et al. 2015).

Specimens examined: CHINA. LIAONING PROVINCE: Benxi, Xiamatang, in a temperate forest with Fagaceae, altitude 430 m, 21 August 2015, Qing Cai 1390 (HKAS 92011); same location, in a temperate forest with Fagaceae, altitude 430 m, 22 August 2015, Qing Cai 1402 (HKAS 92022). SICHUAN PROVINCE: Miyi County, Puwei, altitude 2000 m, 27 July 1986, Ming-Sheng Yuan 1190 (HKAS 18280). JAPAN. HOKKAIDO: Tomakomai, altitude 300 m, 14 September 2011, Zhu L. Yang 5560 (HKAS 71045).

Commentary: Amanita imazekii was described from Japan by Oda et al. (2001), and subsequently reported from China and the Republic of Korea (Yang 2005, 2015; Cho et al. 2015). Phylogenetically, A. imazekii is closely related to A. squarrosipes Zhu L. Yang et al. (Fig. 1b). The differences between the two species are given in the commentary on A. squarrosipes. For detailed descriptions, comparisons with similar species, line drawings and images of A. imazekii see Oda et al. (2001) and Yang (2005, 2015).

34. Amanita incarnatifolia Zhu L. Yang, Bibl. Mycol. 170: 52 (1997).

Replaced name: Amanita rhodophylla Imazeki & Toki, Bull. Govt Forest Exp. Stn Meguro 79: 2 (1955), nom illegit. (a later homonym of Amanita rhodophylla Beeli 1931); non Beeli, Bull. Soc. R. Bot. Belg. 63(2): 103 (1931).

Pileus 3.5–8 cm in diam., grayish (1B1), gray (1C1–3), gray-brown (2C2–4) to brown (1D2–5); volval remnants on pileus absent, or retained as several, irregularly arranged, white (1A1) patches; margin striate (0.2–0.4 R), non-appendiculate. Lamellae pinkish (8A2–3), lamellulae truncate. Stipe 5–10 cm long × 0.5–1.5 cm diam., pinkish (8A2–3) above annulus, white (1A1) under annulus; basal bulb absent; volval remnants on stipe base saccate, white. Annulus subapical, white (1A1) to grayish (1B1).

Basidia 45–65 × 11–16 μm, clavate, 4-spored. Basidiospores [240/13/13] (9.5–) 10.0–13.0 (–16.5) × (6.5–) 7.0–9.0 (–12.0) μm, Q = (1.25–) 1.31–1.5, Qm = 1.4 ± 0.07, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central and southwestern China (Yang 2015; this study), and Japan (this study).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a broad-leaved forest with Fagaceae, altitude 1200 m, 19 July 2017, Yan-Jia Hao 1453 (HKAS 100583); same location, in a forest dominated by Fagaceae, altitude 1200 m, 19 July 2017, Yan-Jia Hao 1470 (HKAS 100593); same location, in a forest dominated by Fagaceae, altitude 1200 m, 19 July 2017, Yan-Jia Hao 1471 (HKAS 100594); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 986 m, 20 July 2017, Yan-Jia Hao 1475 (HKAS 100596); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 1121 m, 21 July 2017, Yan-Jia Hao 1497 (HKAS 100601); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 955 m, 23 July 2017, Yan-Jia Hao 1551 (HKAS 100626); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 955 m, 23 July 2017, Yan-Jia Hao 1552 (HKAS 100627); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 900 m, 25 July 2017, Yan-Jia Hao 1561 (HKAS 100629); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 1200 m, 25 July 2017, Yan-Jia Hao 1571 (HKAS 100634); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 900 m, 25 July 2017, Yan-Jia Hao 1577 (HKAS 100635); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 1200 m, 25 July 2017, Yan-Jia Hao 1590 (HKAS 100637). YUNNAN PROVINCE: Jianchuan County, in a forest of Pinus yunnanensis, altitude 2400 m, 16 August 2003, Zhu L. Yang 4064 (HKAS 43098). JAPAN. AOMORI-KEN: Hakkoda-san, altitude unknown, August 1953, S. Toki (TYPE, TF 3355).

Commentary: Yang (1997) described A. incarnatifolia to replace the name A. rhodophylla Imazeki & Toki, and the basidiospores from the type of this species are [120/4/1] (10.5–) 11.0–14.0 (–16.5) × (7.5–) 8.0–9.5 (–10.5) μm, Q = (1.22–) 1.3–1.56 (–1.69). Yang (2015) then recorded the collection of HKAS 43098 as A. incarnatifolia. In this study, we follow Yang’s treatment and consider the monophyletic clade, which contains the collection of HKAS 43098, as A. incarnatifolia.

Amanita incarnatifolia is phylogenetically related and morphologically similar to A. longistriata S. Imai, A. ristichii Tulloss, A. virginiana (Murrill) Sacc. & Trotter and A. cf. incarnatifolia (Fig. 1b). However, A. longistriata has relatively longer striations on the pileus margin, relatively wider basidiospores (10.0–13.0 × 8.0–11.0 μm, Q = (1.22–) 1.11–1.3, Qm = 1.21 ± 0.08) and is dominant in pine forests (Imai 1938; Gilbert 1940, 1941a; Hongo 1959; Yang and Doi 1999; Yang 2005, 2015; this study). Amanita ristichii has a white basidioma (Tulloss 1989). Amanita virginiana a darker (fuliginous to dark brown) pileus, relatively longer striations on the pileus margin, white to off-white lamellae and relatively wider basidiospores (12.5–13.3 × 9.4–11.7 μm, Q = 1.11–1.45) (Jenkins 1979; Tulloss 1993). Because morphological differences between A. longistriata and A. cf. incarnatifolia (specimens examined listed under A. longistriata) are subtle, more collections are necessary before a formal description of A. cf. incarnatifolia is made. In addition, our multi-locus phylogenetic data indicate that HKAS 29519, treated as A. incarnatifolia in Yang (2005), should be transferred to A. cf. incarnatifolia (Fig. 1b).

Detailed descriptions, comparisons with similar species, line drawings and images of A. incarnatifolia can be found in Yang (2015).

35. Amanita rubromarginata Har. Takah., Mycoscience 45(6): 372 (2004).

Pileus 5–13 cm in diam., plano-convex to applanate, slightly umbonate at center, dark red (9C6–8), red (7B6–8) to orange-red (6B6–8) at center, becoming reddish orange (7A5–8), orange-yellow (5A5–8) to yellow (4A6–8) towards margin; volval remnants on pileus absent; margin striate (ca. 0.5 R), non-appendiculate. Lamellae yellowish (1A2–4); lamellar edges reddish (8A3–5) to orange-red (7A5–8); lamellulae truncate. Stipe 10–22 cm long × 0.8–2.2 cm diam., yellowish (1A2–4) to orange-red (7A5–8), densely covered with orange-red (6B6–8) squamules; basal bulb absent; volval remnants on stipe base saccate, 3.5–6 cm high × 2–2.5 cm wide, outer surface white (1A1) to dirty white (2B1), inner surface cream (1A2) to yellowish (1A2–4). Annulus superior, reddish (8A3–5) to orange-red (6B6–8).

Basidia 25–35 × 9–11 μm, clavate, 4-spored. Basidiospores [80/6/6] 7.5–10.0 (–10.5) × (5.5–) 6.0–7.5 (–8.0) μm, Q = 1.23–1.42 (–1.5), Qm = 1.33 ± 0.07, broadly ellipsoid to ellipsoid, inamyloid. Volval remnants on stipe base composed of two parts intergrading to each other. Outer part of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant; inflated cells scarce. Inner part of volval remnants on stipe base composed of longitudinally arranged to interwoven elements: filamentous hyphae very abundant; inflated cells fairly abundant to locally abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests with Fagaceae (e.g. Castanopsis, Lithocarpus and Quercus); basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (this study), and Japan (Takahashi 2004; Imazeki et al. 2011).

Specimens examined: CHINA. YUNNAN PROVINCE: Mengla County, in a broad-leaved forest with Castanopsis and Lithocarpus, altitude 1130 m, 8 July 2014, Gang Wu 1281 (HKAS 89018); Lancang County, in a subtropical broad-leaved forest with Fagaceae, altitude 1350 m, 31 August 2017, Zhu L. Yang 6051 (HKAS 101240); same location, in a subtropical broad-leaved forest with Fagaceae, altitude unknown, 26 September 2016, LC-LJW 193 (HKAS 97697); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 1200 m, 9 November 2017, LC-LJW 971 (HKAS 101400). JAPAN. OKINAWA-KEN: Ishigaki-shi, Banna-dake, altitude unknown, 15 September 2003, H. Takahashi s.n. (HOLOTYPE, KPM-NC0011979); same location, altitude unknown, 8 June 2002, H. Takahashi s.n. (KPM-NC0010087).

Specimen of A. javanica examined: MALAYSIA. JOHOR STATE: Endau-Rompin National Park, altitude unknown, 21 Mar 2008, S.S. Lee et al. (FRIM 61503).

Specimen of A. cf. javanica examined: CHINA. HAINAN PROVINCE: Ledong County, Jianfengling, in a broad-leaved forest with Fagaceae, altitude 900 m, 5 August 2009, Nian-Kai Zeng 454 (HKAS 59789).

Commentary: Amanita rubromarginata was described from Japan by Takahashi (2004). In our ITS phylogenetic analysis (Fig. S7), collections of TRTC-150314 (ITS = JX844717) and DMSC09996 (ITS = KT213721), which were identified as A. hemibapha s. l. by Sánchez-Ramírez et al. (2015) and Amanita sp. 2 by Parnmen et al. (2016) separately, clustered within A. rubromarginata with strong support. Thus, we consider that these two collections should be assigned to A. rubromarginata.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. rubromarginata is somewhat related to A. javanica. We have checked a specimen of A. javanica from Malaysia. Amanita javanica can be distinguished from A. rubromarginata by its orange-yellow to ocher yellow pileus with a relatively paler reddish tinge, an orange annulus with a less reddish tinge, and its distribution in tropical forests dominated by Dipterocarpaceae (Corner and Bas 1962; Tang et al. 2018; this study). The basidiospores of A. javanica were [20/1/1] 7–9.5 (–10.5) × 5.5–7 (–9) μm, Q = 1.14–1.36, Qm = 1.25 ± 0.06.

HKAS 59789, treated as “A. javanica” in Yang (2015) but indicated in our phylogenetic trees as A. cf. javanica (Fig. 1b), is related to A. rubromarginata and A. javanica. Furthermore, A. cf. javanica has a reddish orange pileus, yellow lamellae, a yellowish stipe densely covered with reddish orange squamules, a reddish orange annulus and basidiospores measuring [100/3/2] 7.5–10.0 × (5.5–) 6.0–7.5 μm, Q = 1.23–1.45 (–1.58), Qm = 1.34 ± 0.07, which are similar to those of A. javanica and A. rubromarginata. Further studies are needed to understand its taxonomy.

36. Amanita kitamagotake N. Endo & A. Yamada, Mycoscience 58: 463 (2017).

Pileus 6–10 cm in diam., yellow (3A4–8) over entire disk; volval remnants on pileus absent; margin striate (0.3–0.4 R), non-appendiculate. Lamellae yellowish (1A2–4); lamellulae truncate. Stipe 8–15 cm long × 1–1.5 cm diam., yellowish (1A2–4) to yellow (3A4–8); basal bulb absent; volval remnants on stipe base saccate, white (1A1). Annulus subapical, yellow (3A4–8).

Basidia 38–56 × 9–12 μm, clavate, 4-spored. Basidiospores [100/7/6] 8.5–10.5 (–11.0) × 6.0–7.5 μm, Q = 1.32–1.47 (–1.57), Qm = 1.41 ± 0.06, ellipsoid, inamyloid. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2015 as Amanita sp. 6; this study) and Japan (Endo et al. 2017).

Specimens examined: CHINA. YUNNAN PROVINCE: Binchuan County, Jizushan, in a subtropical mixed forest with Pinus, Quercus and Rhododendron, altitude 1990 m, 11 August 2011, Li-Ping Tang 1534 [HKAS 69995, Amanita sp. 6 in Yang (2015)]; Nanjian County, in a mixed forest with Fagaceae and Pinaceae, altitude 2300 m, 4 August 2015, Qing Cai 1375 (HKAS 91997); same location, in a mixed forest dominated by Lithocarpus and Castanopsis, altitude 2300 m, 4 August 2015, Qing Cai 1380 (HKAS 92002); Lanping County, in a forest dominated by Pinus, altitude 3200 m, 13 August 2010, Bang Feng 812 (HKAS 68593); Lijiang, Lijiang Alpine Botanic Garden, in a forest with Pinus and Rhododendron, altitude 3350 m, 14 August 2013, Jiao Qin 780 (HKAS 81218); same location, in a forest with Pinus and Rhododendron, altitude ca. 3300 m, 19 August 2013, Jiao Qin 823 (HKAS 81261).

Commentary: Amanita kitamagotake was listed as Amanita sp. 6 from China by Yang (2015) and formally described from Japan by Endo et al. (2017). It is edible and commonly called “Huangluosan” in Yunnan, China.

Species to which A. kitamagotake is phylogenetically related remain as yet unknown (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. kitamagotake see Yang (2015) under Amanita sp. 6.

37. Amanita longistriata S. Imai, Journ. Facul. Agr. Hokkaido Imp. Univ. Sapporo 43: 11 (1938).

Basidiomata slender. Pileus 3–9 cm in diam., grayish (1B1–3), gray (1C1–3) to gray-brown (2C2–5), often brown (3C2–4) at center; volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate. Lamellae pinkish (8A2–4) to pink (9A2–4); lamellulae truncate. Stipe 6–15 cm long × 0.5–1.5 cm diam., white (1A1); basal bulb absent; volval remnants on stipe base saccate, white (1A1). Annulus subapical.

Basidia 35–50 × 11–13 μm, clavate, 4-spored. Basidiospores [160/10/10] (9.5–) 10.0–13.0 (–14.0) × (7.5–) 8.0–11.0 (–12.0) μm, Q = (1.03–) 1.11–1.3 (–1.56), Qm = 1.21 ± 0.08, subglobose to broadly ellipsoid, inamyloid. Clamps present in all parts of basidiomata.

Habitat: Solitary to scattered in pine forests; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2005, 2015; this study), Japan (Imai 1938; Yang and Doi 1999; Imazeki et al. 2011), Republic of Korea (Jo et al. 2013; Jang 2014) and Russian Far East (Malysheva et al. 2014).

Specimens examined: CHINA. YUNNAN PROVINCE: Lanping County, in a forest of Pinus yunnanensis, altitude 2600 m, 15 August 2010, Xue-Tai Zhu 155 (HKAS 68331); Yulong Naxi Autonomous County, in a pine forest, altitude 2000 m, 17 July 2008, Li-Ping Tang 347 (HKAS 54578); same location, in a pine forest, altitude 2488 m, 19 August 2007, Li-Ping Tang 239 (HKAS 53282); same county, in a forest of Pinus yunnanensis, altitude 2600 m, 20 August 2010, Xue-Tai Zhu 255 (HKAS 68431). JAPAN. HONSHU: Yamanashi Prefecture, Mt. Fuji, Yoshida-guchi, in a forest associated with Abies, altitude unknown, 6 September 1983, C. Bas 9040 (L, identified by C. Bas). HOKKAIDO: altitude unknown, 12 August 1935, S. Imai s.n. (TYPE, SAPA); Chiyoda-ku, garden of Fukiage-Gosho Imperial Palace, altitude unknown, 12 July 1995, T. Tobishima s.n. (TNS-F-180370). TOKYO: Oume, Kurosawa 3-chorme, the left-side ridge of Kurosawa River, altitude unknown, 16 July 1997, Y. Doi & K. Miyazaki s.n. (TNS-F-182475); same location, altitude unknown, 16 July 1997, Y. Doi & K. Miyazaki s.n. (TNS-F-182478); same location, altitude unknown, 5 October 1997, Y. Doi s.n. (TNS-F-237593).

Specimens of A. cf. incarnatifolia examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a forest dominated by Fagaceae, altitude 939 m, 19 July 2017, Yan-Jia Hao 1432 (HKAS 100579); same location, in a mixed forest with Fagaceae and Pinaceae, altitude unknown, 25 July 2017, Yan-Jia Hao 1570 (HKAS 100633). HUBEI PROVINCE: Huanren County, in a forest of Fagaceae, altitude 397 m, 26 August 2015, Qing Cai 1429 (HKAS 92048).

Commentary: Amanita longistriata was described from Japan by Imai (1938), and subsequently reported from China, the Republic of Korea and the Russian Far East (Yang 2005, 2015; Jo et al. 2013; Jang 2014; Malysheva et al. 2014). We have examined the type of A. longistriata and the basidiospores are [100/1/1] (9.5–) 10.0–13.0 (–14.0) × (7.5–) 8.0–11.0 (–12.0) μm, Q = (1.03–) 1.11–1.35 (–1.56), Qm = 1.22 ± 0.08. In our multi-locus phylogenetic analysis (Fig. 1b), the collection of C. Bas 9040, which was identified as A. longistriata by C. Bas, clusters with the Chinese ones as a monophyletic clade with good support. In addition, the morphological characters of collections from this clade match well with the protologue, especially the subglobose basidiospores. Thus, we treat this clade as A. longistriata.

Amanita longistriata is phylogenetically related and morphologically similar to A. incarnatifolia, A. ristichii, A. virginiana and A. cf. incarnatifolia (Fig. 1b). However, these species can be distinguished from A. longistriata in the following ways. Amanita incarnatifolia has relatively shorter striations on the pileus margin, relatively narrower basidiospores (10.0–13.0 × 7.0–9.0 μm, Q = 1.31–1.5, Qm = 1.4 ± 0.07) and is dominant in pine, broad-leaved or mixed forests with Pinaceae and Fagaceae (Imazeki and Toki 1955; Yang 1997, 2005, 2015; this study). Amanita ristichii has a white basidioma (Tulloss 1989). Amanita virginiana has a darker (fuliginous to dark brown) pileus and white to off-white lamellae (Jenkins 1979; Tulloss 1993). Amanita cf. incarnatifolia has relatively narrower basidiospores (10.0–13.5 × 8.0–10.0 μm, Q = 1.25–1.5, Qm = 1.36 ± 0.08) and is dominant in broad-leaved or mixed forests, but more collections are needed to allow a formal description of A. cf. incarnatifolia.

Detailed descriptions, comparisons with similar species, line drawings and images of A. longistriata are given in Yang (2005, 2015).

38. Amanita ochracea (Zhu L. Yang) Yang-Yang Cui, Qing Cai & Zhu L. Yang, comb. & stat. nov.

Figures 3i, 10.
Fig. 10

Features of Amanita ochracea (TYPE, HKAS 29522). a Basidiomata; b Lamellar trama, subhymenium and hymenium; c Basidiospores. Bars: a = 3 cm, b–c =  10 μm (Yang 1997)

Mycobank: MB 824971

Basionym: Amanita hemibapha var. ochracea Zhu L. Yang, Bibl. Mycol. 170: 67 (1997).

Type: CHINA. YUNNAN PROVINCE: Lijiang, Heibaishui, in a subalpine forest dominated by Abies, altitude 3200 m, 3 August 1995, Zhu L. Yang 2130 (TYPE, HKAS 29522).

Basidioma (Figs. 3i, 10a) large to very large. Pileus 10–25 cm diam., convex when young, plano-convex to applanate when mature, often umbonate at center, dark brown (5D2–5) when young, but becoming brown (4C3–6) to yellow-brown (3C3–6) at center and yellow-brown (3C3–6) to yellow (2B3–6) at margin when mature; volval remnants on pileus absent; margin striate (0.3–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (1A4–6), occasionally yellow (2B3–6); lamellar edges yellow (4A6–8) to brown (4C3–6); lamellulae truncate. Stipe 15–35 cm long × 2–3 (–5) cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, yellow (4A6–8) to yellowish (3A2–5), with its surface covered with yellow-brown (3C3–6) to red-brown (5B3–6), snakeskin-shaped squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 4–7 (–12) cm high × 3–5 (–7) cm wide, membranous, both surfaces white (1A1). Annulus apical, yellow (4A6–8) at upper surface, and yellow-brown (3C3–6) at lower surface. Odor indistinct.

Lamellar trama bilateral. Mediostratum 20–35 μm wide, composed of elongate to subfusiform inflated cells (60–90 × 15–25 μm); filamentous hyphae abundant, 2–7 (–12) μm wide; vascular hyphae scarce. Lateral stratum composed of fairly abundant to abundant, subfusiform, ellipsoid to elongate inflated cells (95–180 × 15–30 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae fairly abundant to abundant and 3–7 μm wide. Subhymenium (Fig. 10b) 30–40 μm thick, with 2–3 (–4) layers of subglobose, ovoid to ellipsoid cells, 11–25 × 10–15 μm. Basidia (Fig. 10b) 40–55 (–60) × 10–14 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 10c) [190/9/9] (8.0–) 9.0–12.5 (–17.0) × (6.0–) 7.0–9.0 (–10.0) μm, Q = (1.13–) 1.2–1.5 (–1.89); Qm = 1.36 ± 0.1, broadly ellipsoid to ellipsoid, rarely subglobose or elongate, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, ellipsoid to clavate or sphaeropedunculate inflated cells (25–70 × 12–50 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 80–130 μm thick; upper layer (30–60 μm thick) gelatinized, composed of radially to subradially arranged, thin-walled, colorless or sometimes yellowish brown, filamentous hyphae 2–5 μm wide; lower layer (50–70 μm thick) composed of radially to subradially arranged, filamentous hyphae 3–7 μm wide, colorless or yellowish to brownish; vascular hyphae scarce. Interior of volval remnants on stipe base dominantly composed of longitudinally arranged elements: filamentous hyphae abundant to dominant, 2–8 (–12) μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to locally abundant, becoming more abundant toward inner surface, subglobose, ovoid to ellipsoid, 50–90 × 45–60 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of the volval remnants on stipe base dominantly composed of longitudinally arranged, very abundant to dominant filamentous hyphae, inflated cells scarce and not easily observed; inner surface gelatinized, similar to structure of inner part, but with more filamentous hyphae. Stipe trama composed of longitudinally arranged, long clavate terminal cells, 200–400 × 25–55 μm; filamentous hyphae scattered to abundant, 2–7 μm wide; vascular hyphae scarce. Annulus composed of subradially arranged elements: inflated cells abundant in upper surface, but becoming scarce to scattered toward inner surface, ovoid, subglobose, clavate or sphaeropedunculate, 25–100 × 20–50 μm, colorless, thin-walled; filamentous hyphae fairly abundant to abundant, 2–7 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in subalpine forests of Abies, Picea, Pinus, and Quercus; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 2005, 2015; this study), and India (Upadhyay et al. 2008).

Additional specimens examined: CHINA. TIBET AUTONOMOUS REGION: Linzhi, Lulang, in a subalpine forest, altitude 2900 m, 1 August 2014, Kuan Zhao 668 (HKAS 89599). YUNNAN PROVINCE: Lijiang, Sandaowan, in a subalpine forest of Quercus, altitude 3200 m, 1 August 1995, Zhu L. Yang 2114 (HKAS 29497); Yulong Naxi Autonomous County, in a subalpine forest of Quercus and Pinus, altitude 2950 m, 12 August 2014, Qi Zhao 2129 (HKAS 87895); same location, in a subalpine forest, altitude 3100 m, 4 August 2005, Yan-Chun Li 315 (HKAS 48548); Shangri-La, Xiaozhongdian, in a subalpine forest dominated by Abies and Picea, altitude 3300 m, 21 August 2014, Qi Zhao 2175 (HKAS 87941); same location, in a subalpine forest of Abies and Picea, altitude 3300 m, 21 August 2014, Qi Zhao 2199 (HKAS 87965); same location, in a forest with Picea and Abies, altitude 3460 m, 22 August 2014, Qi Zhao 2207 (HKAS 87973); same location, in a subalpine forest of Abies and Picea, altitude 3300 m, 21 August 2014, Qi Zhao 2220 (HKAS 87986).

Commentary: Amanita ochracea was described as a variety of A. hemibapha based exclusively on morphological data (Yang 1997). In our phylogenetic analysis (Fig. 1b), samples of A. hemibapha var. ochracea Zhu L. Yang and A. hemibapha var. hemibapha cluster in distinct clades. Amanita hemibapha var. ochracea differs from A. hemibapha var. hemibapha by its larger and more robust basidioma with a brown umbonate pileus, and its subalpine distribution. Consequently, we treated A. hemibapha var. ochracea as a novel species, A. ochracea.

In our phylogenetic analyses (Figs. 1b, S7), three collections, RET 317-5 in the multi-locus data, RET 718-7 (ITS = KX810029) and RET 290-5 (ITS = JX 844699) in the ITS data, which were submitted to NCBI as a provisional name of A. cinnamomescens Tulloss et al., clustered together in the monophyletic A ochracea clade. Furthermore, the morphological and ecological differences between these three species and A. ochracea are only subtle and they should therefore be considered A. ochracea. In addition, one specimen, DAK-M-08 (ITS = KC797155), which was submitted to NCBI under the name A. calyptroderma G. F. Atk. & V. G. Ballen, also clustered within the clade of A. ochracea in our ITS analysis (Fig. S7) and should be A. ochracea.

Amanita ochracea is phylogenetically (Figs. 1b, S7) related to A. vernicoccora Bojantchev & R. M. Davis. However, A. vernicoccora, described from the USA, has a relatively smaller basidioma, a yellow pileus covered with a monolithic patch at the center, and ellipsoid to elongate basidiospores (9.2–11.8 × 6.2–7.1 μm) (Bojantchev et al. 2011).

Amanita ochracea is also somewhat similar to A. javanica and A. rubromarginata in its macroscopic appearance. However, the latter two species have a relatively smaller basidioma, an orange-red to red pileus that is 5–13 cm in diameter, yellow lamellae with reddish to orange-red lamellar edges and a reddish to orange-red annulus (Boedijn 1951; Corner and Bas 1962; Takahashi 2004; Imazeki et al. 2011; Lee 2017; Tang et al. 2018). Amanita ochracea is also comparable to the North American A. arkansana H. R. Rosen, but the latter has relatively longer striations on the pileal margin (0.4–0.5 R) and smaller basidiospores (7.5–10.0 × 7.0–8.0 μm) (Rosen 1926; Gilbert 1941a; Jenkins 1986).

39. Amanita princeps Corner & Bas, Persoonia 2(3): 297 (1962).

Pileus 10–20 cm in diam., brownish (2B2–4), brown (1C4–6) to yellow-brown (3C3–6), sometimes becoming brownish (2B2–4) to yellowish (3A2–4) towards margin; volval remnants on pileus absent, or sometimes retained as white (1A1) patches appressed at center; margin shortly striate (0.25–0.3 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 9–25 (–30) cm long × 1–3 cm diam., white (1A1) to dirty white (2B1), with its surface covered with concolorous pulverulent, floccose to fibrous squamules; basal bulb absent; volval remnants on stipe base saccate, outer surface white (1A1), dirty white (2B1) to brownish (3B3–5), often cracking and peeling into thin patches, inner surface dirty white (2B1) to brownish (2B2–2). Annulus apical, white (1A1). Odor indistinct.

Basidia 40–60 × 12–16 μm, clavate, 4-spored. Basidiospores [240/12/7] (7.0–) 8.0–11.0 (–13.0) × (7.0–) 7.5–10.0 (–12.0) μm, Q = 1.0–1.17 (–1.29), Qm = 1.08 ± 0.05, globose, subglobose to broadly ellipsoid, inamyloid. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 3–12 μm wide; inflated cells scarce to locally abundant, subglobose, subfusiform to ellipsoid, 60–90 × 15–65 μm; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical forests with Fagaceae, Pinaceae and Dipterocarpaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China (Yang 2005, 2015; this study), India (Bhatt et al. 2017), Malaysia (Lee 2017; Tang et al. 2018), Singapore (Corner and Bas 1962) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. HAINAN PROVINCE: Ledong County, Jianfengling, in a tropical to subtropical forest, altitude 900 m, 20 July 2009, Xing-Liang Wu 1829 (HKAS 59777); same location, in a tropical to subtropical forest, altitude ca. 900 m, 27 July 2009, Xing-Liang Wu 8220 (HKAS 59779); same location, in a tropical to subtropical forest, altitude ca. 900 m, 28 July 2009, Xing-Liang Wu 1021 (HKAS 59782); same location, in a tropical to subtropical forest dominated by Fagaceae, altitude 900 m, 4 August 2009, Nian-Kai Zeng 434 (HKAS 59793); Qiongzhong County, Limushan, in a tropical to subtropical forest dominated by Fagaceae, altitude 800 m, 3 August 2010, Zai-Wei Ge 2593 [HKAS 60269, as Amanita sp. 8 in Yang (2015)]. MALAYSIA. JOHOR: in a lowland, dipterocarp forest, altitude unknown, 2 September 2009, S.S. Lee et al. (FRIM 62849). KUALA LUMPUR: Kepong, altitude unknown, 6 October 1999, Farid 2830 (FRIM 2830).

Commentary: Amanita princeps was described from Singapore by Corner and Bas (1962), and subsequently reported from China, India, Malaysia and Thailand (Yang 2005, 2015; Sanmee et al. 2008; Bhatt et al. 2017; Lee 2017; Tang et al. 2018). Little information was known of this species from or near its type locality until Tang et al. (2018). Tang et al. (2018) identified A. princeps with both morphological and molecular evidence based on materials from Malaysia (FRIM 62849 and FRIM 2811), which were collected near its type locality and which were morphologically consistent with the original description of A. princeps in Corner and Bas (1962).

In our multi-locus phylogenetic data (Fig. 2), the Chinese specimens clustered with A. princeps from Malaysia (FRIM 62849) as two subclades based on PTP analysis. We have examined a specimen of A. princeps from Malaysia (FRIM 62849), and the basidiospores were [20/1/1] 8.5–10.5 (–11.5) × 7.0–10.0 (–11.5) μm, Q = 1.0–1.18 (–1.29), Qm = 1.09 ± 0.06. There are subtle morphological differences between specimens in the two subclades from China and Malaysia, but they share a large basidioma, a saccate volva on the stipe base with its outer layer often cracking and peeling into thin patches, and globose, subglobose to broadly ellipsoid basidiospores, all of which are consistent with the original description of A. princeps in Corner and Bas (1962). Consequently, we currently conservatively treated the specimens in these two subclades as A. princeps and submitted their DNA sequences to NCBI under that name. In addition, our phylogenetic data indicate that HKAS 60269, treated as Amanita sp. 8 in Yang (2015), should be A. princeps (Figs. 1b, 2).

In our multi-locus phylogenetic analysis (Fig. 1b), A. princeps was closely related to the Australian A. egregia D. A. Reid. However, A. egregia has a white basidioma and has an Australian distribution in association with eucalypt trees (Reid 1978, 1980).

Amanita princeps can be easily confused with A. pseudoprinceps Yang-Yang Cui et al. because of the yellow-brown pileus with short striations (0.25–0.3 R). However, A. pseudoprinceps, a novel species described in this study, differs from A. princeps by its relatively smaller basidioma with pileus ca. 7–12 cm diam., and a white saccate volva with its outer surface not cracking into brownish patches.

Amanita princeps is similar to A. aporema, but the latter has a relatively smaller basidioma with pileus ca. 6–10 cm in diam., a more red-brown pileus with longer striations ca. 0.5 R, and a saccate volva on the stipe base with its outer surface white and not cracking into patches (Boedijn 1951; Lee 2017; Tang et al. 2018; this study).

Detailed descriptions, comparisons with similar species and images of A. princeps are given in Yang (2005, 2015).

40. Amanita pseudoprinceps Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 3j, 11.
Fig. 11

Microscopic features of Amanita pseudoprinceps (TYPE, HKAS 97523). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base; d longitudinal section of inner part of volval remnants on stipe base. Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 824972

Etymology: pseudoprinceps, from pseudo- = false-, and princeps = Amanita princeps, is proposed due to its similarity to A. princeps.

Type: CHINA. YUNNAN PROVINCE: Lancang County, in a mixed forest with Fagaceae and Pinaceae, altitude 1000 m, 19 August 2016, LC-LJW 19 (TYPE, HKAS 97523, GenBank Acc. No.: ITS = MH508527, nrLSU = MH486788, tef1-α = , rpb2 = MH486202, β-tubulin  = MH485710).

Basidioma (Fig. 3j) medium-sized to large. Pileus 7–12 cm diam., convex to applanate, sometimes slightly depressed at center, mostly yellow-brown (3B2–6) to brown (1B2–5) over disk, with olivaceous (30A5–8) tinge, occasionally dark brown (4E5–8) to brown (1B2–5) at center, but becoming cream (1A2–4) to white (1A1) towards margin; volval remnants absent; margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (1A2–4); lamellulae truncate, plentiful. Stipe 10–15 cm long × 1–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with minute, dirty white (2B1) to brownish (2B2–4) fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 4–6 cm high × 3–4 cm wide, membranous, outer surface white (1A1) to dirty white (2B1), inner surface brownish (2B2–4). Annulus apical, white (1A1) to cream (1A2–4), membranous. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–60 μm wide, composed of abundant ellipsoid to clavate inflated cells (60–200 × 10–35 μm); filamentous hyphae abundant, 3–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant clavate inflated cells (25–70 × 12–25 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 11a) 30–60 μm thick, with 2–3 layers of subglobose, fusiform, ellipsoid to irregularly arranged cells, 8–45 × 8–30 μm. Basidia (Fig. 11a) 45–70 × 12–16 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa clamped. Basidiospores (Fig. 11b) [120/6/4] (9.5–) 10.0–12.0 (–14.0) × (8.0–) 8.5–10.5 (–12.0) μm, Q = 1.02–1.23 (–1.27), Qm = 1.12 ± 0.04, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, ellipsoid to fusiform or sphaeropedunculate inflated cells (20–40 × 15–35 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 110–180 μm thick; upper layer (25–100 μm thick) gelatinized, composed of radially arranged to interwoven, thin-walled, colorless, filamentous hyphae 2–5 μm wide; lower layer (60–105 μm thick) composed of radially arranged, filamentous hyphae 2–8 μm wide, colorless to yellowish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 11c, d) dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant, 5–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to fairly abundant, subglobose, subfusiform to ellipsoid, 40–100 × 20–60 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of the volval remnants on stipe base similar to structure of interior part, but with more abundant inflated cells (inflated cells becoming fairly abundant to scattered toward inner part); inner surface gelatinized and similar to structure of interior part. Stipe trama composed of longitudinally arranged, ellipsoid, clavate to long clavate terminal cells, 60–300 × 15–60 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Annulus dominantly composed of two parts intergrading to each other. Outer part of annulus dominantly composed of radially arranged elements: inflated cells scarce to scattered, globose, subglobose to ellipsoid, 15–35 × 12–30 μm, colorless, thin-walled; filamentous hyphae very abundant to dominant, 2–10 μm wide, colorless, thin-walled; vascular hyphae scarce. Inner part of annulus dominantly composed of radially arranged elements: inflated cells fairly abundant to abundant, subglobose, fusiform, ellipsoid to clavate, 20–70 × 10–30 μm, colorless, yellow to yellow-brown, thin-walled; filamentous hyphae abundant, 2–9 μm wide, colorless, yellow to yellow-brown, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China, Nepal and Thailand.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lancang County, in a mixed forest with Fagaceae and Pinaceae, altitude 1000 m, 19 August 2016, LC-LJW 16 (HKAS 97520); same location, in a mixed forest with Fagaceae and Pinaceae, altitude 1400 m, 24 September 2016, LC-LJW 138 (HKAS 97642); Nanjian County, Wuliangshan, in a mixed forest with Fagaceae and Pinaceae, altitude 1600 m, 4 August 2015, Qing Cai 1373 (HKAS 91995).

Specimens of A. aporema examined: INDONESIA. SUMATRA: Batang Paleopoeh, July 1929, E. Jacobson s.n. (HOLOTYPE, BO). MALAYSIA. KUALA LUMPUR: Kepong, FRIM, in a lowland forest, altitude unknown, 1 April 2008, S.S. Lee et al. (FRI 61295). JOHOR: Endau-Rompin, in a lowland forest, altitude unknown, 18 March 2009, S.S. Lee et al. (FRI 62681); same location, in a lowland forest, altitude unknown, 18 March 2009, S.S. Lee et al. (FRI 62674).

Commentary: Amanita pseudoprinceps is characterized by its yellow-brown to brown pileus with shorter striations on the margin (0.2–0.3 R), an apical annulus, subglobose to broadly ellipsoid basidiospores (10–12 × 8.5–10.5 μm) and the common presence of clamps.

In our ITS phylogenetic analysis (Fig. S7), three specimens, DMSC10886 (ITS = KT213715) and DMSC10883 (ITS = KT213710) from Thailand, and CMU-MC06 (ITS = JX844763) from Nepal, clustered within A. pseudoprinceps with strong support (MLB = 100%). Thus, these three specimens should be A. pseudoprinceps.

Our multi-locus phylogenetic analysis (Fig. 1b) indicated that A. pseudoprinceps was sister to A. aporema Boedijn. Based on the original description and the examinations of collections, A. aporema differs from A. pseudoprinceps by its relatively smaller basidioma with pileus ca. 6–10 cm in diam., and a relatively darker colored pileus (red-brown) with relatively longer striations (ca. 0.5 R) (Boedijn 1951; Lee 2017; Tang et al. 2018; this study).

Amanita pseudoprinceps can be confused with A. princeps as they share a yellow-brown pileus with short striations (0.2–0.3 R). However, A. princeps differs from A. pseudoprinceps in its relatively larger basidioma with pileus ca. 10–20 cm in diam., and a white saccate volva with its outer surface cracking and peeling into thin, brownish patches (Corner and Bas 1962; Yang 2015; Lee 2017; Tang et al. 2018).

Amanita pseudoprinceps is similar to the African A. zambiana Pegler & Piearce in appearance, but the latter has a relatively larger basidioma with pileus ca. 10–20 cm, relatively shorter striations on the pileal margin (0.1 R), saccate volval remnants on the stipe base with its outer surface cracking into large, indefinite verrucae, outer surface blackish brown, inner surface red-brown (Pegler and Piearce 1980; Pegler and Shah-Smith 1997).

Amanita malayensis Tang et al. and A. similis Boedijn are also similar to A. pseudoprinceps. However, A. malayensis has a yellow pileus with its center often yellowish orange, maize yellow or saffron yellow and ellipsoid basidiospores (9.0–12.0 × 6.0–8.0 μm, Q = 1.28–1.57, Qm = 1.39 ± 0.13) (Tang et al. 2018). Amanita similis has relatively longer striations on the pileal margin (0.4–0.6 R), a bright yellow stipe with its surface marked with pinkish orange scales, a pinkish orange to grayish annulus, and relatively smaller basidiospores (8.0–9.0 × 6.0–7.0 μm, Q = 1.23–1.41, Qm = 1.3 ± 0.06) (Boedijn 1951; Corner and Bas 1962; Lee 2017; Tang et al. 2018).

41. Amanita rubroflava Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 3k, l, 12.
Fig. 12

Microscopic features of Amanita rubroflava. a Hymenium and subhymenium (TYPE, HKAS 83089); b basidiospores (TYPE, HKAS 83089); c longitudinal section of outer part of volval remnants on stipe base (HKAS 83715); d longitudinal section of inner part of volval remnants on stipe base (HKAS 83715). Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 824973

Etymology: rubroflava, from rubro- = red-, flava = yellow, referring to its red center and yellow margin of pileus.

Type: CHINA. YUNNAN PROVINCE: Nanjian County, Lingbaoshan, in a broad-leaved forest dominated by Fagaceae, altitude 2300 m, 3 August 2014, Yan-Jia Hao 1298 (TYPE, HKAS 83089, GenBank Acc. No.: ITS = MH508568, nrLSU = MH486827, tef1-α  = MH509054, rpb2 = MH486238, β-tubulin  = MH485743).

Basidioma (Fig. 3k, l) medium-sized to large. Pileus 7–12 cm diam., convex to applanate, distinctly umbonate, red (8C8, 9B8) to orange-red (7B7–8) at center, but becoming yellow (5B6–8, 4A6–8) to yellowish (2A4–6) at margin; volval remnants on pileus absent; margin striate (0.2–0.4 R), non-appendiculate; trama white, yellowish to brown, unchanging. Lamellae free, crowded, yellow (1A4–7); lamellulae truncate, plentiful. Stipe 15–22 cm long × 0.7–3 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, yellow (4A6–8, 1A4–7) and covered with concolorous, irregularly arranged, snakeskin-shaped squamules; context white (1A1) to yellowish (1A2–4), hollow in center; basal bulb absent; volva saccate, 5–7 cm high × 3–5 cm wide, membranous, both surfaces white (1A1) to dirty white (2B1). Annulus apical to subapical, yellow (4A6–8) to yellowish (2A3–5). Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–60 μm wide, composed of abundant ellipsoid, fusiform to clavate inflated cells (55–110 × 15–35 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid, fusiform to clavate, inflated cells (35–120 × 20–35 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–10 μm wide. Subhymenium (Fig. 12a) 40–50 μm thick, with 2–3 layers of subglobose, ellipsoid to fusiform or irregular cells, 10–25 (–45) × 10–20 μm. Basidia (Fig. 12a) 35–55 × 9–13 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 12b) [210/7/7] (7.0–) 8.0–10.0 (–11.0) × (6.0–) 6.5–8.5 (–9.0) μm, Q = 1.05–1.31 (–1.4), Qm = 1.18 ± 0.06, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (10–30 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–10 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 60–150 μm thick; upper layer (15–40 μm thick) slightly or non-gelatinized, composed of radially to interwoven, thin-walled, colorless filamentous hyphae 2–5 (–15) μm wide; lower layer (70–125 μm thick) composed of radially arranged filamentous hyphae 2–8 μm wide, colorless to yellow brown; vascular hyphae scarce. Volval remnants on stipe base (Fig. 12c, d) composed up of two parts intergrading to each other. Outer part (Fig. 12c) composed up of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 3–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, subfusiform to ellipsoid, 45–90 × 20–50 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Inner part (Fig. 12d) composed up of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to locally abundant, subglobose, fusiform to ellipsoid, 45–110 × 20–65 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volva dominantly composed of very abundant filamentous hyphae, occasionally mixed with scarce inflated cells; inner surface gelatinized, similar to structure of inner part, but with more abundant filamentous hyphae. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 50–300 × 15–40 μm; filamentous hyphae scattered to abundant, 2–9 μm wide; vascular hyphae scarce. Annulus composed of subradially arranged elements: inflated cells scarce to scattered, subglobose, ellipsoid, fusiform to clavate, 30–70 × 15–30 μm, colorless, thin-walled; filamentous hyphae very abundant to nearly dominant, 2–10 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Dali, Jizushan, in a broad-leaved forest dominated by Fagaceae, altitude 2600 m, 8 August 2014, Qing Cai 1279 (HKAS 83748); Nanjian County, Ailaoshan, in a broad-leaved forest dominated by Fagaceae, altitude 2580 m, 6 August 2014, Qing Cai 1246 (HKAS 83715); Nanjian County, Lingbaoshan, in a broad-leaved forest dominated by Fagaceae, altitude 2350 m, 3 August 2014, Qing Cai 1189 (HKAS 83649); Jingdong County, Ailaoshan, in a subtropical broad-leaved forest, altitude 2500 m, 20 July 2006, Zhu L. Yang 4677 [HKAS 50474, as Amanita sp. 4 in Yang (2015)]; same location, in a forest dominated by Fagaceae, altitude 2300 m, 23 July 2013, Bang Feng 1361 (HKAS 101434); Yulong Naxi Autonomous County, in a mixed forest with Fagaceae and Pinaceae, altitude 2600 m, 31 August 2013, Bang Feng 1470 (HKAS 82576).

Commentary: Amanita rubroflava is characterized by a umbonate pileus, which is red to orange-red at the center, but yellow and long-striate at the margin, subglobose to broadly ellipsoid basidiospores (8.0–10.0 × 6.5–8.5 μm), and the common presence of clamps.

Our ITS phylogenetic analysis (Fig. S7) suggests that A. rubroflava is closely related to A. aff. caesaroides. The latter was identified based on the collection RET-356-10 (ITS = MF928093) by Sánchez-Ramírez et al. (2015). Whether RET-356-10 represents A. rubroflava or a new taxon needs further morphological studies with more collections.

In China, A. rubroflava can be confused with A. hemibapha (Zhang et al. 2004; Yang 2005). Our ITS phylogenetic analysis (Fig. S7) showed that the collection HKAS 38416 (ITS = AY436460), identified as A. hemibapha in Zhang et al. (2004) and Yang (2005), clustered within the A. rubroflava clade with strong support. Consequently, HKAS 38416 should be determined as A. rubroflava. In addition, A. hemibapha differs morphologically from A. rubroflava by its non-umbonate pileus, narrower basidiospores (8.0–10.0 × 5.5–6.5) and its tropical distribution (Gilbert 1941a; Pegler 1986; Vrinda et al. 2005; Sanmee et al. 2008; Yang 2015).

Amanita rubroflava is similar to A. caesareoides in that they share umbonate pilei and the basidiospores are of similar size, however, the latter has a more red to orange-red pileus and is dominant in temperate East Asia (Imazeki and Hongo 1987; Imazeki et al. 1988; Cho et al. 2015; Yang 2015; Endo et al. 2016).

Amanita caesarea and A. masasiensis Härk. & Saarim. are also comparable to A. rubroflava in their macroscopic appearance. However, the European A. caesarea has a robuster basidioma with an obviously shorter stipe, an orange pileus lacking an umbo and longer basidiospores (8.5–12.0 × 6.0–8.0 μm) (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; this study). Amanita masasiensis, described from Tanzania, has a relatively smaller basidioma, a non-umbonate pileus, ellipsoid to elongate basidiospores (9.0–11.0 × 6.0–7.0 μm), and is dominant in woodland often with trees of the genera Brachystegia, Combretum, Julbernardia, Syzygium and Uapaca (Härkönen et al. 1994; Tang et al. 2015).

42. Amanita similis Boedijn, Sydowia 5(3-6): 322 (1951).

Pileus dark brown (4E4–6), brown (4D4–6) to olivaceous brown (3C3–6); volval remnants on pileus absent; margin striate. Lamellae yellowish (2A2). Stipe covered with orange-red (7A5–7) to yellowish (3A2–5) squamules. Annulus orange-red (7A5–7) to yellow (4A5–7).

Habitat: Solitary to scattered in tropical to subtropical forests of broad-leaved trees; basidioma occurring in summer and autumn.

Distribution: Known from central and eastern China (Yang 2005; this study), Japan (Imazeki et al. 2011) and tropical, southeastern Asia regions (Boedijn 1951; Corner and Bas 1962; Lee 2017; Tang et al. 2018).

Specimens examined: CHINA. HUNAN PROVINCE: Ling County, in a subtropical forest, altitude unknown, 24 July 1990, collector unknown (MHHNU 4519). TAIWAN PROVINCE: Nantou County, Lianhuachi, in a tropical to subtropical forest, altitude 750 m, 5 June 2002, Jian-Ming Chen 3180 (HKAS 41321); Pingdong County, Chufengshan, in a tropical to subtropical forest, altitude 300 m, 13 May 1998, Wen-Neng Chou 03187 (TNM-F0010997); same location, in a tropical to subtropical forest, altitude 300 m, 13 May 1998, Weng-Neng Chou 03188 (TNM-F0010998). MALAYSIA. NEGERI SEMBILAN: Pasoh, in a lowland forest, altitude unknown, 6 September 2006, D. Stubbe 06106 (FRI 3740). JOHOR: in a lowland dipterocarp forest, altitude unknown, 5 September 09, Lee et al. (FRI 69312).

Commentary: We examined two collections of A. similis from Malaysia (FRI 3740 and 69312). The basidiospores were [40/2/2] (7.5–) 8.0–10.0 × 6.0–7.5 μm, Q = 1.21–1.47 (–1.67), Qm = 1.34 ± 0.09, broadly ellipsoid to ellipsoid. Yang (2005) cited four specimens under the name A. similis, which were morphologically similar to A. similis, but no DNA sequences were generated from the collections. In this study, we continue to treat these four Chinese specimens as A. similis.

Our multi-locus phylogenetic data (Fig. 1b) indicate that A. similis is related to A. subhemibapha and A. fuscoflava. However, A. subhemibapha has a yellow to yellowish pileus with its center orange and relatively shorter striations (0.25–0.35 R) on the pileal margin. Amanita fuscoflava has a yellow stipe decorated with yellow squamules and a yellow annulus.

Morphologically, A. similis is similar to A. malayensis Tang et al., A. princeps Corner and Bas and A. aporema Boedijn. However, these species differ from A. similis in the following ways. A. malayensis has a yellow pileus with its center yellowish orange, maize yellow or saffron yellow, relatively shorter striations (0.3–0.4 R) on the pileal margin, a yellow stipe decorated with yellow scales, and a yellowish white to whitish annulus (Tang et al. 2018). Amanita princeps has a relatively larger basidioma with pileus ca. 10–20 cm, relatively shorter striations on the pileal margin (0.25–0.3 R), a white stipe, a white annulus, a saccate volva with its outer surface cracking into patches, and relatively broader basidiospores (8.0–11.0 × 7.5–10.0 μm, Q = 1.0–1.17, Qm = 1.08 ± 0.05) (Corner and Bas 1962; Yang 2015; Lee 2017; Tang et al. 2018; this study). Amanita aporema has a dirty white to cream stipe, a whitish to cream annulus, and relatively broader basidiospores (9.5–11.0 × 8.5–10.5 μm, Q = 1.02–1.11, Qm = 1.06 ± 0.04) (Boedijn 1951; Lee 2017; Tang et al. 2018; this study).

Amanita chatamagotake N. Endo & A. Yamada and A. hayalyuy D. Arora & G. H. Shepard are comparable to A. similis. However, A. chatamagotake has a yellow or grayish yellow stipe without orange scales, a yellow annulus and relatively shorter striations (0.2–0.5 R) on the pileal margin (Endo et al. 2017). Amanita hayalyuy has a relatively larger basidioma with pileus ca. 8–18 cm, a relatively brighter colored pileus (golden-brown or bright yellow-brown), relatively shorter striations on pileal margin, a yellowish annulus and relatively larger basidiospores (9.3–11.7 × 6.2–7.8 μm) (Shepard et al. 2008).

43. Amanita squarrosipes Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 3m, n, 13.
Fig. 13

Microscopic features of Amanita squarrosipes (TYPE, HKAS 76359). a Hymenium and subhymenium; b basidiospores. Bars: ab = 10 μm

MycoBank: MB 825003

Etymology: squarrosipes, from squarrosipes = squamules on stipe, referring to its stipe covered with gray squamules.

Type: CHINA. SICHUAN PROVINCE: Tibetan Autonomous County of Muli, in a subalpine forest with Picea, Quercus and Rhododendron, altitude 3500 m, 1 August 2012, Yan-Jia Hao 701 [TYPE, HKAS 76359, as Amanita cf. imazekii in Yang (2015), GenBank Acc. No.: ITS = MH508613, nrLSU = MH486894, tef1-α  = MH509116, rpb2 = MH486297, β-tubulin  = MH485797].

Basidioma (Fig. 3m, n) medium-sized. Pileus ca. 6.5 cm diam., convex to applanate, lacking an umbo at center, pale grayish (1B1, 2B1, 3B1) to brownish gray (2B2–4) over disk; volval remnants on pileus absent; margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (2A2–3); lamellulae truncate, plentiful. Stipe ca. 19 cm long × 1.3 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to pale grayish (1B1, 2B1, 3B1), glabrous above annulus, and densely covered with gray (4C1–2) to dark gray (3E1–4) squamules under annulus; context white (1A1), hollow in center; basal bulb absent; volva saccate, ca. 5 cm high × 2.5 cm wide, membranous, both surfaces white (1A1) to dirty white (2B1). Annulus subapical, pendant from attachment ca. 3 cm below apex of stipe, membranous, upper surface pale grayish (1B1, 2B1, 3B1), lower surface gray brownish (2B2–4). Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant ellipsoid, fusiform to clavate inflated cells (60–120 × 15–40 μm); filamentous hyphae abundant, 2–13 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid to fusiform inflated cells (40–55 × 15–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 13a) 30–50 μm thick, with 2–3 layers of subglobose, ellipsoid to irregular cells, 10–25 × 10–20 μm. Basidia (Fig. 13a) 50–80 × 12–17 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 13b) [40/1/1] 9.0–11.0 (–11.5) × (8.0–) 8.5–10.5 μm, Q = 1.0–1.1, Qm = 1.05 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells [10–40 (–80) × 10–35 μm], single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–5 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 90–125 μm thick; upper layer (25–50 μm thick) slightly gelatinized, composed of radially arranged, thin-walled, colorless, filamentous hyphae 2–5 μm wide; lower layer (50–90 μm thick) composed of radially arranged, filamentous hyphae 2–10 μm wide, colorless to brownish; vascular hyphae scarce. Volval remnants on stipe base dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to fairly abundant, subglobose, fusiform to ellipsoid, 35–100 × 20–80 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Stipe trama composed of longitudinally arranged, long clavate terminal cells (25–) 70–350 × 15–40 (–60) μm; filamentous hyphae scattered to abundant, 2–12 μm wide; vascular hyphae scarce. Annulus composed of radially arranged elements: inflated cells scarce, ellipsoid to cylindrical, 30–100 × 10–20 μm, colorless, thin-walled; filamentous hyphae very abundant to dominant, 2–10 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: On soil in a subalpine forest with Picea, Quercus and Rhododendron.

Distribution: Known from southwestern China.

Commentary: Amanita squarrosipes is well distinguished by its pale gray pileus, a white stipe densely covered with gray to dark gray squamules, globose to subglobose basidiospores (9.0–11.0 × 8.5–10.5 μm), the common presence of clamps and distributions in subalpine forests.

This taxon was phylogenetically sister to and morphologically similar to A. imazekii T. Oda, C. Tanaka & Tsuda (Fig. 1b). However, A. imazekii, a species described from Japan and also found in China, has a relatively larger basidioma, and a white stipe densely covered with white to grayish squamules (Oda et al. 2001; Yang 2005, 2015).

Morphologically, A. squarrosipes is similar to A. murrilliana Singer, A. esculenta Hongo & I. Matsuda, A. princeps Corner & Bas and A. longistriata S. Imai. However, A. murrilliana has a white stipe and relatively broader basidiospores (7.8–8.6 × 11.7–13.3 μm) (Murrill 1941; Singer 1951; Jenkins 1979). Amanita esculenta has relatively shorter striations (0.1 R) on the pileal margin and relatively longer basidiospores (10.0–12.0 × 7.5–9.0 μm, Q = 1.24–1.45, Qm = 1.35 ± 0.09) (Matsuda and Hongo 1955; Imazeki and Hongo 1987; Yang 2005, 2015; Imazeki et al. 2011). Amanita princeps has a relatively larger basidioma with a pileus ca. 10–20 cm in diam., a white stipe covered with concolorous squamules, a saccate volva with its outer surface often cracking into pieces and is distributed in tropical to subtropical forests (Corner and Bas 1962; Yang 2015; Lee 2017; Tang et al. 2018). Amanita longistriata has relatively longer striations (0.3–0.5 R) on the pileal margin, pinkish to pink lamellae, a white stipe and relatively longer basidiospores (10.0–13.0 × 8.0–11.0 μm, Q = 1.11–1.3, Qm = 1.21 ± 0.08) (Imai 1938; Gilbert 1940, 1941a; Hongo 1959; Yang and Doi 1999; Yang 2005, 2015; this study).

44. Amanita subhemibapha Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 3o, 14.
Fig. 14

Microscopic features of Amanita subhemibapha. a Hymenium and subhymenium (TYPE, HKAS 96847); b basidiospores (TYPE, HKAS 96847); c longitudinal section of interior part of volval remnants on stipe base (HKAS 97518). Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825004

Etymology: subhemibapha, from sub- = near, hemibapha = Amanita hemibapha, referring to its similarity to A. hemibapha.

Type: CHINA. YUNNAN PROVINCE: Lancang County, in a subtropical mixed forest with Fagaceae and Pinaceae, altitude 1300 m, 19 August 2016, LC-Hao 10 (TYPE, HKAS 96847, GenBank Acc. No.: nrLSU = MH486906, tef1-α  = MH509127, rpb2 = MH486307).

Basidioma (Fig. 3o) medium-sized to large. Pileus 5–11 cm diam., convex, plano-convex to applanate, lacking an umbo at center, purely orange (5B5–8) when young, but becoming orange (5B5–8) at center and yellow (4A6–8) to yellowish (3A3–6) at margin when mature; volval remnants on pileus absent; margin striate (0.25–0.35 R), non-appendiculate; trama white (1A1), but yellow (4A6–8) to yellowish (3A3–6) at the lower part of pileipellis, unchanging. Lamellae free, crowded, white (1A1) to cream (1A4–6); lamellar edges yellow (4A6–8); lamellulae truncate, plentiful. Stipe 5–16 cm long × 0.8–1.7 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, yellow (4A6–8) to orange (5B5–8), with its surface covered with concolorous, snakeskin-shaped squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 3.5–6 cm high × 3–4 cm wide, membranous, both surfaces white (1A1). Annulus apical to subapical, yellow (4A6–8) to orange (5B5–8). Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–75 μm wide, composed of abundant ellipsoid, fusiform to clavate inflated cells (35–80 × 10–25 μm); filamentous hyphae abundant, 2–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid to fusiform inflated cells (20–85 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 3–8 μm wide. Subhymenium (Fig. 14a) 30–50 μm thick, with 2–3 layers of subglobose to ellipsoid cells, 10–25 × 8–20 μm. Basidia (Fig. 14a) 40–55 × 9–12 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 14b) [180/9/3] (7.0–) 8.0–11.0 × (5.5–) 6.0–8.0 (–9.0) μm, Q = 1.15–1.53 (–1.67), Qm = 1.34 ± 0.07, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (8–50 × 8–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–4 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 90–180 μm thick; upper layer (35–150 μm thick) gelatinized, composed of radially to interwoven, thin-walled, colorless, filamentous hyphae 2–5 μm wide; lower layer (30–60 μm thick) composed of radially arranged, filamentous hyphae 3–8 (–10) μm wide, colorless; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 14c) dominantly composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, fusiform to ellipsoid, 55–100 × 20–70 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of the volval remnants on stipe base similar to structure of interior part, but with more abundant filamentous hyphae; inner surface gelatinized, similar to structure of inner part. Stipe trama composed of longitudinally arranged, long clavate terminal cells, 60–250 × 20–60 μm; filamentous hyphae scattered to abundant, 2–9 (–15) μm wide; vascular hyphae scarce. Annulus composed of subradially arranged to interwoven elements: inflated cells scarce, subglobose, fusiform to clavate, 20–100 × 10–35 μm, colorless, thin-walled; filamentous hyphae very abundant to dominant, 2–10 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China and Thailand.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lancang County, in a subtropical forest dominated by Fagaceae, altitude 1000 m, 26 September 2016, LC-LJW 193 (HKAS 97697); same location, in a subtropical mixed forest with Fagaceae and Pinaceae, altitude 1000 m, 19 August 2016, LC-LJW 14 (HKAS 97518).

Commentary: Amanita subhemibapha is characterized by its non-umbonate orange pileus, white to cream lamellae with yellow edges, a yellow to orange stipe, broadly ellipsoid to ellipsoid basidiospores (8.0–11.0 × 6.0–8.0 μm) and the common presence of clamps.

In our phylogenetic analysis (Fig. 1b), A. subhemibapha was closely related to A. fuscoflava. For the comparisons between the two species see the commentary on A. fuscoflava. In our ITS phylogenetic analysis (Fig. S7), collection TRTC-150286 (ITS = JX844714) from Thailand, which was identified as A. hemibapha s. l. by Sánchez-Ramírez et al. (2015), clustered with A. subhemibapha and should be a renamed as such.

Species morphologically comparable to A. subhemibapha include A. hemibapha, A. javanica and A. rubromarginata. However, A. hemibapha possesses a redder pileus and narrower basidiospores (8.0–10.0 × 5.5–6.5) (Gilbert 1941a; Pegler 1986; Vrinda et al. 2005; Sanmee et al. 2008; Yang 2015; this study). Amanita javanica and A. rubromarginata have a redder pileus with relatively longer striations on the pileal margin (ca. 0.5 R), yellow lamellae with reddish to orange-red lamellar edges and a reddish to orange-red annulus (Boedijn 1951; Corner and Bas 1962; Takahashi 2004; Imazeki et al. 2011; Lee 2017; Tang et al. 2018; this study).

Amanita rubroflava Yang-Yang Cui, Qing Cai & Zhu L. Yang, A. caesarea and A. caesareoides are also similar to A. subhemibapha in appearance. However, A. rubroflava, a novel species described in this study, differs from A. subhemibapha in its distinctly umbonate pileus and slightly broader basidiospores (8.0–10.0 × 6.5–8.5 μm). The European A. caesarea has a robuster basidioma with a shorter stipe, a redder pileus with shorter striations on its margin and larger basidiospores (8.5–12.0 × 6.0–8.0 μm) (Gilbert 1940, 1941a; Contu 2000a; Floriani 2000; Wasser 2000; Galli 2001; Neville and Poumarat 2004; this study). Amanita caesareoides has a redder umbonate pileus, and is dominant in temperate East Asia (Imazeki and Hongo 1987; Imazeki et al. 1988; Cho et al. 2015; Yang 2015; Endo et al. 2016).

45. Amanita yuaniana Zhu L. Yang, Mycotaxon 51: 464 (1994).

Pileus 7–13 cm in diam., gray (1B1–2), gray-brown (1C2–5) to brown (4C2–4), with innate spots or speckles; volval remnants on pileus absent; margin striate (up to 0.1 R), non-striate. Lamellae white (1A1); lamellulae truncate. Stipe 7–14 cm long × 1–2.5 cm diam., white (1A1) to grayish (1B1); basal bulb absent; volval remnants on stipe base saccate, white (1A1) to grayish (1B1). Annulus subapical, white (1A1) to grayish (1B1).

Basidia 48–75 × 10–13 μm, clavate, 4-spored. Basidiospores [390/13/8] (8.5–) 9.5–12.0 (–13.0) × (6.0–) 6.5–8.0 (–9.5) μm, Q = (1.27–) 1.31–1.61 (–1.74), Qm = 1.45 ± 0.09, ellipsoid, inamyloid. Interior of volval remnants on stipe base dominantly composed of very abundant to dominant filamentous hyphae. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in pine or mixed forests with Fagaceae and Pinaceae at altitude ca. 2000–3000 m; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1994, 1997, 2005, 2015; this study).

Specimens examined: CHINA. SICHUAN PROVINCE: Miyi County, altitude 2000 m, 27 July 1986, Ming-Sheng Yuan 1187 (HOLOTYPE, HKAS 18383). YUNNAN PROVINCE: Jianchuan County, in a forest of Pinus, altitude 2260 m, 29 August 2009, Qing Cai 170 (HKAS 58835); Lijiang, Xuesongcun, in a forest of Pinus, altitude 2700 m, 27 August 2009, Qing Cai 142 (HKAS 58807); Lijiang, Yulongxueshan, in a forest of Pinus, altitude 2600 m, 17 August 2010, Bang Feng 881 (HKAS 68662); Yulong Naxi Autonomous County, in a forest of Pinus, altitude 2550 m, 1 September 2009, Qing Cai 206 (HKAS 59971); same county, in a forest dominated by Pinaceae, altitude 2000 m, 17 July 2008, Li-Ping Tang 346 (HKAS 54577); same county, Yulongxueshan, in a forest of Pinus yunnanensis and P. armandii, altitude ca. 2300 m, 31 July 1995, Zhu L. Yang 2103 (HKAS 29516); same county, Shitouxiang, in a forest of Pinus yunnanensis, altitude 2550 m, 1 September 2009, Qing Cai 206 (HKAS 58871).

Commentary: Amanita yuaniana was described from China by Yang (1994). Our phylogenetic analysis (Fig. 1b) indicates that A. yuaniana is related to A. hunanensis. However, A. hunanensis differs from A. yuaniana in the relatively longer striations on its margin (ca. 0.2–0.3 R) and fairly abundant inflated cells in the volval remnants on the stipe base (Peng and Liu 1981; Yang and Zhang 2002; Yang 2005, 2015).

Amanita esculenta and A. spreta are also morphologically similar to A. yuaniana. However, A. esculenta can be distinguished from A. yuaniana by its pileus lacking innate speckles (Matsuda and Hongo 1955; Imazeki and Hongo 1987; Yang 2005, 2015; Imazeki et al. 2011). Amanita spreta, described form the USA, has a pileus lacking innate speckles and relatively narrower basidiospores (10.2–13.3 × 5.5–7 μm) (Murrill 1913; Coker 1917; Jenkins 1978, 1986; Tulloss et al. 1995). For detailed descriptions, comparisons with similar species, line drawings and images of A. yuaniana see Yang (1994, 1997, 2005, 2015).

Amanita sect. Vaginatae (Fr.) Quél., Mém. Soc. Emul. Montbéliard, Ser. II, 5: 64 (1872).

Basionym: Agaricus sect. Vaginatae Fr., Monogr. Amanit. Sueciea: 2 (1854).

Lectotype: Amanita vaginata (Bull.: Fr.) Lam., Encycl. Méth. Bot. (Paris) 1(1): 109 (1783).

Diagnosis: Basidioma agaricoid; pileus striate and non-appendiculate; volval remnants on pileus usually absent; stipe base bulbless, with saccate volva or warts arranged in incomplete belts; annulus mostly absent, but occasionally clearly present; clamps absent in all parts of basidioma.

Commentary: The recognition and characterization of the species in Amanita sect. Vaginatae is often difficult (Bas 1977). In this study, we documented 32 Chinese taxa from this section based on multi-locus phylogenetic, morphological and partial ecological evidence, including 13 novel species and 19 known taxa (17 species and two varieties). Amanita atrofusca Zhu L. Yang was treated as a novel synonym of A. umbrinolutea (Gillet) Bataille. The microscopic structure of the volva, including the arrangement and proportion of inflated cells and filamentous hyphae, seems to be one of the most important traits for the characterization of the species in this section. In addition, DNA sequences are very useful for delimitation of the species.

Key to the species of Amanita sect. Vaginatae from China

  1. 1.

    Annulus absent……….2

     
  2. 1.

    Annulus present………A. cingulata

     
  3. 2.

    Volval remnants on pileus often absent, or occasionally retained as white patches; volval remnants on stipe base membranous and saccate……….3

     
  4. 2.

    Volval remnants on pileus present, verrucose, conical to felted; volval remnants on stipe base verrucose, subpyramidal to conical, often arranged in incomplete belts on lower part of stipe……….29

     
  5. 3.

    Lamellae white, cream to grayish; stipe white, dirty white, grayish to brownish……….4

     
  6. 3.

    Lamellae red to pink; stipe pinkish………A. pallidocarnea

     
  7. 4.

    Outer surface of volval remnants on stipe base coarse, with verrucose or tomentose to felted, brown to brownish squamules……….5

     
  8. 4.

    Outer surface of volval remnants on stipe base glabrous, lacking verrucae……….6

     
  9. 5.

    Pileus ca. 5.5 cm in diam., brown to grayish, with a dark ring-like zone at proximal end of marginal striations; stipe white, covered with snakeskin-shaped, gray squamules; outer surface of volval remnants on stipe base covered with felted to tomentose or verrucose, brown squamules; basidiospores 11.5–14.0 × 11.0–13.5 μm………A. tomentosivolva

     
  10. 5.

    Pileus 7–14 cm in diam., orange-red at center, orange-yellow to yellow at margin, lacking a ring-like zone; stipe white to cream, covered with snakeskin-shaped, yellow to orange-red squamules; outer surface of volval remnants on stipe base covered with verrucose, brown to brownish squamules; basidiospores 10.0–12.5 × 9.0–12.0 μm………A. verrucosivolva

     
  11. 6.

    Basidiospores globose to subglobose, Q = 1.0–1.15……….7

     
  12. 6.

    Basidiospores subglobose to broadly ellipsoid, or broadly ellipsoid to ellipsoid, Q = 1.05–1.51……….23

     
  13. 7.

    Basidioma not white……….8

     
  14. 7.

    Basidioma white………A. vaginata var. alba

     
  15. 8.

    Pileus often with an distinctly dark-colored ring-like zone at proximal end of marginal striations……….9

     
  16. 8.

    Pileus often lacking an distinctly dark-colored ring-like zone at proximal end of marginal striations……….11

     
  17. 9.

    Basidiospores relatively larger, 10.0–13.0 × 9.0–12.0 μm; basidioma distributed in alpine to subalpine forests……….10

     
  18. 9.

    Basidiospores relatively smaller, 9.0–10.5 × 8.5–10.0 μm; basidioma distributed in subtropical broad-leaved forests………A. zonata

     
  19. 10.

    Pileus relatively darker, gray, gray-brown to dark gray; stipe white to dirty white, covered with gray, gray-brown to dark gray squamules………A. umbrinolutea

     
  20. 10.

    Pileus relatively lighter, brown to brownish; stipe white to brownish, covered with concolorous squamules………A. lignitincta

     
  21. 11.

    Pileus often with obvious umbo at center……….12

     
  22. 11.

    Pileus often lacking obvious umbo at center……….16

     
  23. 12.

    Basidioma slender; pileus gray-brown, gray to brown, lacking obvious yellow tinge……….13

     
  24. 12.

    Basidioma robust; pileus yellow-brown to yellow at center, but yellow to yellowish toward margin, with obvious yellow tinge………A. orienticrocea

     
  25. 13.

    Basidioma distributed in temperate mixed forest with Fagaceae, Pinaceae and Betulaceae (European countries and northeastern China) or in subalpine forest associated with Abies, Betula, Pinus and Quercus (southeastern China)……….14

     
  26. 13.

    Basidioma distributed in broad-leaved forests with Fagaceae (central China)………A. shenongjiana

     
  27. 14.

    Pileus often dark brown at center, becoming brown to brownish toward margin; volval remnants on pileus mostly absent……….15

     
  28. 14.

    Pileus often gray-brown to gray over the entire disk; volval remnants on pileus absent or occasionally retained as white patches………A. griseoumbonata

     
  29. 15.

    Basidioma relatively slender with stipe ca. 15–22.5 cm long; lower part of volval remnants on stipe base often not tightly appressed on stipe………A. changtuia

     
  30. 15.

    Basidioma relatively robust with stipe ca. 10–15 cm long; lower part of volval remnants on stipe base often tightly appressed on stipe………A. battarrae

     
  31. 16.

    Pileal margin with long striations, ca. 0.5 R; volval remnants on stipe base microscopically not separated into two parts, inflated cells distributed evenly; basidioma distributed in broad-leaved forests……….17

     
  32. 16.

    Pileal margin with short striations, 0.1–0.3 R; volval remnants on stipe base microscopically not or separated into two parts, and inflated cells distributed evenly, or concentrated in outer or inner part; basidioma dominant in pine, broad-leaved or mixed forests……….18

     
  33. 17.

    Pileus gray to gray-brown; basidiospores relatively smaller, (9.0–) 9.5–11.0 (–12.0) × (8.0–) 9.5–10.5 (–11.5) µm………A. angustilamellata

     
  34. 17.

    Pileus brown to yellow-brown; basidiospores relatively larger, 10.0–12.0 (–12.5) × (9.0–) 10.0–11.5 (–12.5) μm………A. tenuifulva

     
  35. 18.

    Pileus dark gray, gray-brown, gray, grayish to dirty white……….19

     
  36. 18.

    Pileus brown to yellow-brown…………….22

     
  37. 19.

    Basidioma distributed in areas near-snow line and with Polygonum, Potentilla, Rhododendron, Cyperus and Juniperus………A. nivalis

     
  38. 19.

    Basidioma distributed in areas not near snow-line……….20

     
  39. 20.

    Pileus dark gray to gray-brown; stipe dirty white covered with dark gray fibrils………A. brunneoprocera

     
  40. 20.

    Pileus gray-brown to gray; stipe white to dirty white without distinct squamules……….21

     
  41. 21.

    Volval remnants often persistent as large, white patches slightly attached on pileus; the upper part of volva on the stipe base often easily cracking into large patches slightly attached on lower part of stipe………A. retenta

     
  42. 21.

    Volval remnants often absent; volva on the stipe base not easily broken………A. vaginata var. vaginata

     
  43. 22.

    Basidioma relatively larger with pileus ca. 5–15 cm in diam.; basidiospores relatively larger, (8.5–) 10.0–14.0 (–17.0) × (8.0–) 9.5–13.0 (–16.5) μm; inflated cells in volval remnants on stipe base dominantly concentrated in inner part; basidioma distributed in subalpine forests………A. orientifulva

     
  44. 22.

    Basidioma relatively smaller with pileus ca. 6–8 cm diam.; basidiospores relatively smaller, (8.5–) 9.0–11.0 (–13.5) × (7.5–) 8.0–10.0 (–13.0) μm; inflated cells in volval remnants on stipe base scarce to fairly abundant and distributed evenly; basidioma distributed under pine forests of Pinus yunnanensis………A. albidostipes

     
  45. 23.

    Pileus dark brown, gray-brown, gray to brown; stipe white, gray to brown……….24

     
  46. 23.

    Pileus brown at center, but yellow at margin; stipe yellow, often with pale pinkish color at lower part………A. hamadae

     
  47. 24.

    Basidiospores broadly ellipsoid to ellipsoid, Q = 1.15–1.51……….25

     
  48. 24.

    Basidiospores subglobose to broadly ellipsoid, occasionally globose, Q = 1.05–1.25……….27

     
  49. 25.

    Pileus gray to grayish, lacking olivaceous tinge……….26

     
  50. 25.

    Pileus brown, with obviously olivaceous tinge………A. olivaceofusca

     
  51. 26.

    Pileus 4–7 cm in diam., glabrous, margin with long striations (0.3–0.4 R); volva on stipe base large; basidiospores 9.0–11.0 × 7.0–9.0 μm, Q = 1.13–1.38………A. ovalispora

     
  52. 26.

    Pileus 3–6 cm in diam., often with felted to verrucose, grayish volval remnants, margin with short striations (0.2–0.3 R); volva on stipe base small; basidiospores 9.5–12.5 × 8.0–10.5 μm, Q = 1.05–1.33………A. pseudovaginata

     
  53. 27.

    Basidioma relatively smaller with pileus 3.5–9 cm diam.; pileus relatively paler, yellow-brown; basidioma dominant in pine, broad-leaved or mixed forests……….28

     
  54. 27.

    Basidioma relatively larger with pileus 5–14 cm diam.; pileus dark brown to gray-brown; basidioma dominant in subalpine forests………A. brunneofuliginea

     
  55. 28.

    Pileus with an indistinctly dark-colored ring-like zone at the proximal end of marginal striations; volval remnants on stipe base microscopically not separated into two parts, and inflated cells distributed evenly………A. pallidozonata

     
  56. 28.

    Pileus lacking an indistinctly dark-colored ring-like zone at proximal end of marginal striations; volval remnants on stipe base microscopically separated into two parts, and inflated cells concentrated in outer part………A. chiui

     
  57. 29.

    Basidiospores globose to subglobose……….30

     
  58. 29.

    Basidiospores subglobose to broadly ellipsoid………A. neocinctipes

     
  59. 30.

    Basidiospores relatively larger, mostly more than 11 μm in diam……….31

     
  60. 30.

    Basidiospores relatively smaller, mostly less than 11 μm in diam………A. cinctipes

     
  61. 31.

    Pileus 10–14 cm in diam., dark brown to black, covered with felted, verrucose to irregular, dark gray to brown-gray squamules; stipe robust, 1.5–3 cm wide, covered with gray to black squamules; basidiospores globose to subglobose, (11.0–) 11.5–15.0 (–24.0) × (9.5–) 11.0–14.5 (–20.0) μm; basidioma distributed in alpine to subalpine pine forests………A. liquii

     
  62. 31.

    Pileus 3–7 cm in diam., gray to brown-gray, covered with removable, pulverulent, conical to felted squamules; stipe slender, 0.5–1.5 cm wide, covered with gray squamules; basidiospores globose to subglobose, (9.5–) 10.0–13.5 (–16.5) × (8.5–) 9.5–13 (–15) μm; basidiospores distributed in subtropical pine, broad-leaved or mixed forests………A. griseofolia

     

46. Amanita albidostipes Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 15a, 16.
Fig. 15

Fresh basidiomata of novel species in Amanita sect. Vaginatae. a A. albidostipes (TYPE, HKAS 57358); b–c A. changtuia (TYPE, HKAS 92100); d–f A. chiui (de TYPE, HKAS 76328, f HKAS 54613); g–i A. griseoumbonata (g–h TYPE, HKAS 92103, i HKAS 75594); j–l A. lignitincta (j–k HKAS 69411, l HKAS 58788); m–o A. neocinctipes (m TYPE, HKAS 79627, no HKAS 78463). Bars: 2 cm

MycoBank: MB 825028

Etymology: albidostipes, from albidus = whitish, and stipes = stipe, referring to the whitish to dirty white stipe.

Type: CHINA. YUNNAN PROVINCE: Changning County, in a forest dominated by Pinus yunnanensis, altitude 1500 m, 20 August 2009, Bang Feng 629 (TYPE, HKAS 57358, GenBank Acc. No.: ITS = MH508500, nrLSU = MH486756, tef1-α  = MH508983).

Basidioma (Fig. 15a) medium-sized. Pileus 6–8 cm diam., plano-convex to applanate, yellowish (1A2–4), yellow-brown (4B2–5) to brown (3B3–5), occasionally whitish (1A1), dirty white (1B1) to brownish (2B2–4); volval remnants on pileus usually absent; margin striate (0.1–0.25 R), non-appendiculate; trama white (1A1) to dirty white (2B1), unchanging. Lamellae free, crowded, white (1A1) to cream (2A2); lamellulae truncate, plentiful. Stipe 6.5–11 cm long × 0.9–2 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with concolorous, slightly recurved fibrils; context white (1A1) to dirty white (2B1), hollow in center; basal bulb absent; volva saccate, 2–3.5 cm high × 1.2–2 cm wide, membranous, outer surface white (1A1) to dirty white (2B1) and often with brown (3B3–5) tinge, inner surface dirty white (2B1), grayish (1B1–2) to grayish brown (2B2–4). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–60 (–75) μm wide, composed of abundant subglobose, fusiform, ellipsoid to clavate inflated cells (30–100 × 15–40 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid to fusiform inflated cells (25–60 × 8–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 16a) 30–45 μm thick, with 2–3 layers of ellipsoid to fusiform or irregularly arranged cells, 25–40 (–60) × 8–30 μm. Basidia (Fig. 16a) 40–55 × 10–16 μm, clavate, 4-spored; sterigmata 3–7 μm long; basal septa lacking clamps. Basidiospores (Fig. 16b) [140/5/2] (8.5–) 9.0–11.0 (–13.5) × (7.5–) 8.0–10.0 (–13.0) μm, Q = 1.00–1.15 (–1.22), Qm = 1.08 ± 0.05, globose to subglobose, rarely broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of ellipsoid to fusiform or sphaeropedunculate inflated cells (10–60 × 10–45 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 100–175 μm thick; upper layer (40–100 μm thick) gelatinized, composed of interwoven to radially arranged, thin-walled, colorless, filamentous hyphae 2–6 μm wide; lower layer (25–85 μm thick) composed of radially arranged, filamentous hyphae 2–8 (–10) μm wide, colorless to brownish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 16c) composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to fairly abundant, subglobose, ellipsoid to clavate, 45–110 × 15–70 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer and inner surfaces of volval remnants on stipe base similar to structure of interior part, but with inner surface gelatinized. Stipe trama composed of longitudinally arranged, clavate terminal cells, 50–300 × 15–55 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.
Fig. 16

Microscopic features of Amanita albidostipes (TYPE, HKAS 57358). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm

Habitat: Solitary to scattered on soil under pine forests of Pinus yunnanensis; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimen examined: CHINA. YUNNAN PROVINCE: Qiubei County, in a forest of Pinus yunnanensis, altitude 1557 m, 9 August 2014, Pan-Meng Wang 108 (HKAS 95189).

Commentary: Amanita albidostipes is characterized by its yellow-brown to brown pileus, a white to whitish stipe, globose to subglobose basidiospores (9.0–11.0 × 8.0–10.0 μm) and association with trees of the genus Pinus. It is worth mentioning that specimens have also been collected with whitish, dirty white to brownish pilei, which may be caused by mutations or differently environmental conditions.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. albidostipes is related to A. brunneoprocera and A. orienticrocea. However, A. brunneoprocera can be easily distinguished from A. albidostipes by its dark gray, gray-brown to gray pileus and a white to dirty white stipe covered with gray to dark gray fibrils. For comparisons between A. albidostipes and A. orienticrocea see the commentary on A. orienticrocea.

The European A. crocea is similar to A. albidostipes in the color of basidiomata and the size of basidiospores, but the former differs by its brighter colored basidioma with an orange tinge, and its stipe surface densely covered with orange to orange-ocheraceous squamules (Gilbert 1940, 1941a; Contu 2000a, b; Massart 2000; Tulloss 1997, 2000a; Galli 2001).

The yellow-brown pileus and globose to subglobose basidiospores of A. orientifulva are also comparable to those of A. albidostipes, but A. orientifulva has a larger basidioma, larger basidiospores (10.0–14.0 × 9.5–13.0 μm), more inflated cells in the inner part of the volva on the stipe base, and is found in subalpine forests (Yang 2005, 2015).

Amanita hamadae, a species described from Japan, is similar to A. albidostipes in morphological appearance. However, A. hamadae has broadly ellipsoid to ellipsoid basidiospores (10.0–12.0 × 7.5–9.0 μm) and is dominant in subalpine forests with trees of the genera Picea and Abies (Nagasawa and Hongo 1984; Yang 2015).

47. Amanita angustilamellata (Höhn.) Boedijn, Sydowia 5(3–6): 318 (1951).

Basionym: Amanitopsis vaginata var. angustilamellata Höhn., Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1 123: 74 (1914).

Basidioma slender. Pileus 4–7 cm in diam., depressed at center, gray-brown (4E3–7) to gray (1D1–3); volval remnants on pileus absent; margin striate (ca. 0.5 R), non-striate. Lamellae sparse, white (1A1); lamellulae truncate. Stipe 8–12 cm long × 0.5–1.5 cm diam., dirty white (2B1), grayish (2C1–2) to brownish (2D2–4); basal bulb absent; volval remnants on stipe base saccate, small, 2–3 cm high × 1–2 cm wide. Annulus absent.

Basidia 46–66 × 12–17 μm, clavate, 4-spored. Basidiospores [30/4/4] (9.0–) 9.5–11.0 (–12.0) × (8.0–) 9.5–10.5 (–11.5) μm, Q = 1.0–1.05 (–1.13), Qm = 1.03 ± 0.13, globose to subglobose, inamyloid. Interior of volval remnants on stipe base composed of irregularly arranged elements: filamentous hyphae abundant; inflated cells fairly abundant to abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from northern and southwestern China (Yang 2005; this study), Indonesia (Boedijn 1951), Malaysia (Lee 2017) and Singapore (Corner and Bas 1962).

Specimens examined: CHINA. Hainan Province: Ledong County, Jianfengling, 30 September 1987, Thai-Hui Li (GDGM 12578); Qiongzhong County, Limushan, altitude 800–850 m, 24 August 1988, Guo-Yang Zheng (GDGM 14315). Yunnan Province, Jiangcheng, Hongjiang, altitude 1350 m, 6 August 1991, Pei-Gui Liu 851 (HKAS 24158). INDONESIA. JAVA: Tjibodas, 1908, Fr. von Höhnel s.n. (TYPE, FH 4712).

Commentary: Amanita angustilamellata was described from Java by Boedijn (1951), and subsequently reported from China, Malaysia and Singapore (Corner and Bas 1962; Yang 2015; Lee 2017). This species is characterized by its gray-brown pileus without an umbo, longer striations up to 0.5 R on the pileal margin, sparse lamellae, a small, white saccate volva on stipe base, globose to subglobose basidiospores (9.5–11.0 × 9.5–10.5 μm, Q = 1.0–1.05, Qm = 1.03 ± 0.13), and the absence of clamps.

The diagnosis mentioned in the protologue of this taxon (as Amanitopsis vaginata var. angustilamellata) was very brief, and since then few reliable descriptions have been made (Boedijn 1951; Corner and Bas 1962; Pegler 1986; Yang 2005, 2015; Lee 2017). Consequently, the delimitation of A. angustilamellata remains poorly understand.

Corner and Bas (1962) mentioned that the narrow gills of this species may be an unreliable character. Pegler (1986) reported the presence of clamps in the pileipellis based on Sri Lankan materials, but this differed from the reports of Corner and Bas (1962) and Yang (2005). Boedijn (1951) and Lee (2017) did not mention the presence or absence of clamps of this species. Based on the protologue and reexamination of the type (FH 4712), HKAS 24158 was identified as A. angustilamellata by Weiß et al. (1998) and Yang (2005). In this study, we follow the treatments of A. angustilamellata in Weiß et al. (1998) and Yang (2005). For detailed descriptions, comparisons with similar species, line drawings and images of the Chinese A. angustilamellata see Yang (2005).

Yang (2015) reported the presence of A. angustilamellata in China based on HKAS 83453. However, our multi-locus phylogenetic analysis (Fig. 1b) indicated that HKAS 83453 grouped together with HKAS 89451 as a distinct clade, and collections in this clade had a more northern subtropical to temperate distribution compared with A. angustilamellata. Thus, HKAS 83453 and HKAS 89451 may represent a novel species, but its classification cannot be clarified until further collections are made to better estimate morphological characters and geographic distributions.

48. Amanita battarrae (Boud.) Bon, Docums Mycol. 16 (no. 61): 16 (1985).

Pileus 3–6 cm in diam., convex-applanate to applanate, more or less umbonate at center, dark brown (4E4–6) to gray-brown (3E1–4) at center, becoming grayish brown (2D3–5), brown (2C4–7) to brownish (2B2–4) toward margin; volval remnants on pileus absent; margin striate (0.25–0.4 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate; lamellar edge white (1A1). Stipe 10–15 cm long × 0.5–1.5 cm diam., white (1A1) to dirty white (2B1), covered with white (1A1), dirty white (2B1) to brownish (1C2–3) fibers; basal bulb absent; volval remnants on stipe base saccate, ca. 3–4 cm in height, with lower part often tightly appressed to stipe surface, both surfaces white (1A1), outer surface occasionally with yellow-brown (4B3–5) spots. Annulus absent.

Basidia 48–60 × 13–18 μm, clavate, 4-spored. Basidiospores [150/9/9] (9.0–) 9.5–12.0 (–14.0) × (8.5–) 9.0–10.5 (–13.5) μm, Q = 1.0–1.08 (–1.12), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Clamps absent in all parts of basidiomata.

Habit: Solitary to scattered in forests with Abies, Betula, Pinus and Quercus; basidioma occurring in summer and autumn.

Distribution: Known from European countries (Consiglio 2000; Neville and Poumarat 2009), northeastern and southwestern China (this study), the Islamic Republic of Iran (Bahram et al. 2006) and Russian Far East (Malysheva and Kovalenko 2015).

Specimens examined: CHINA. JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest with Pinaceae, altitude 1160 m, 4 August 2008, Yan-Chun Li 1415 (HKAS 56255); same location, in a forest with Abies, Betula and Pinus, altitude 1162 m, 30 August 2015, Qing Cai 1464 (HKAS 92083); same location, in a forest with Abies, Betula and Pinus, altitude 1162 m, Qing Cai 1472 (HKAS 92090). SICHUAN PROVINCE: Kangding County, in a mixed forest, altitude 3500 m, 7 September 2016, KD-Ding 68 (HKAS 101399); Tibetan Autonomous County of Muli, in a mixed forest with Fagaceae and Pinaceae, altitude 2500 m, 4 September 2011,Yan-Jia Hao 590 (HKAS 76348). TIBET AUTONOMOUS REGION: Changdu, in a subalpine to alpine forest, altitude 3200 m, 25 July 2009, Bang Feng 500 (HKAS 57229); Gonjo County, in a subalpine to alpine forest, altitude 3500 m, 31 July 2009, Bang Feng 534 (HKAS 57263). YUNNAN PROVINCE: Lijiang, in a mixed forest dominated by Pinus yunnanensis, altitude 2700 m, 3 August 2011, Qing Cai 532 (HKAS 70128). AUSTRIA. VORARLBERG: Kleinwalsertal, Hirschegg, in a forest with Picea and Abies, altitude 1150 m, 5 September 2011, KR 10641 (MB-000643).

Commentary: Amanita battarrae is characterized by a gray-brown pileus with its center more or less umbonate, a white saccate volva on the stipe base with its lower part often tightly appressed to the stipe surface, the outer surface of the volva occasionally with yellow-brown spots, globose to subglobose basidiospores (9.5–12.0 × 9.0–10.5 μm) and an association with trees of the genera Abies, Betula, Pinus and Quercus.

In our multi-locus phylogenetic data (Fig. 2), collections labeled as A. battarrae could be split into two closely related phylogenetic species based on GCPSR analysis. Since few morphological differences were observed between these two phylogenetic species, we treat them as a single species in this study, a conclusion which was also supported by our PTP analysis (Fig. 2). No species closely related to A. battarrae were detected from our current data (Fig. 1b). Furthermore, our phylogenetic analysis (Fig. 1b) suggested that HKAS 56255, treated as A. vaginata (Bull.) Lam. var. vaginata by Yang (2015), should be A. battarrae.

Amanita battarrae is morphologically similar to A. brunneofuliginea Zhu L. Yang, A. umbrinolutea (Gillet) Bataille and A. griseoumbonata Yang-Yang Cui et al. However, A. brunneofuliginea has a relatively larger basidioma with a pileus ca. 5–14 cm in diam. and relatively broader basidiospores (10.5–13.0 × 9.5–12.0 μm, Q = 1.06–1.24, Qm = 1.13 ± 0.06) (Yang 1997, 2005, 2015). Amanita umbrinolutea has a relatively darker colored pileus and stipe, and brownish to brown lamellar edges (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015; this study). Amanita griseoumbonata has a saccate volva with its lower part not tightly appressed to the stipe surface, and its outer surface is white without yellow pots.

49. Amanita brunneofuliginea Zhu L. Yang, Bibl. Mycol. 170: 96 (1997).

Pileus 5–14 cm in diam., umbonate at center, dark brown (3F4–8) to brown (3E6–8) or gray-brown (3E2–5); volval remnants on pileus absent; margin striate (0.2–0.4 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 8–22 cm long × 0.5–3 cm diam., white (1A1) to dirty white (2B1); basal bulb absent; volval remnants on stipe base saccate, 2.5–7 cm high × 1–3.5 cm wide, with its outer surface covered with brown (3E6–8) patches. Annulus absent.

Basidia 46–74 × 15–23 μm, clavate, 4-spored. Basidiospores [200/20/19] (10.0–) 10.5–13.0 (–14.0) × (9.0–) 9.5–12.0 (–12.5) μm, Q = (1.04) 1.06–1.24 (–1.26), Qm = 1.13 ± 0.06, subglobose to broadly ellipsoid, inamyloid. Brown patches on outer surface of volval remnants on stipe base dominantly composed of very abundant to dominant filamentous hyphae and scarce inflated cells. Volval remnants on stipe base dominantly composed of two layers intergrading to each other. Outer layer composed of irregularly arranged, abundant filamentous hyphae and very abundant inflated cells. Inner layer composed of longitudinally arranged, very abundant filamentous hyphae mixed with locally abundant inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subalpine forests with Abies, Picea, Quercus and Pinus; basidioma occurring in summer and autumn.

Distribution: Known from central, northern, northeastern, northwestern and southwestern China (Yang 1997, 2005, 2015; this study).

Specimens examined: CHINA. GANSU PROVINCE: Zhouqu County, in a subalpine forest of Abies, altitude 2400 m, 16 August 2012, Xue-Tai Zhu 718 (HKAS 76567). HEILONGJIANG PROVINCE: Yichun, in a forest with Abies, Pinus koraiensisa and Betula platyphylla, altitude 470 m, 3 September 2015, Qing Cai 1492 (HKAS 92110); same location, in a forest with Pinus koraiensisa and Betula platyphylla, altitude 470 m, 3 September 2015, Qing Cai 1495 (HKAS 92078). SICHUAN PROVINCE: Dawu County, in a subalpine forest, altitude 3544 m, 17 July 2014, Kuan Zhao 563 (HKAS 89226); same location, in a subalpine forest, altitude 3544 m, 17 July 2014, Jian-Wei Liu 84 (HKAS 90878); same location, in a subalpine forest, altitude 3600 m, 17 July 2014, Bang Feng 1596 (HKAS 93980); Dege County, Yulonglacuo, in a subalpine forest, altitude 3600 m, 9 August 2013, Kuan Zhao 336 (HKAS 80761); Hongyuan County, in a subalpine forest, altitude 3563 m, 19 July 2014, Kuan Zhao 574 (HKAS 89503); same location, in a subalpine forest, altitude 3563 m, 19 July 2014, Jian-Wei Liu 116 (HKAS 90910); Kangding County, in a subalpine forest, altitude 3000 m, 5 September 2016, KD-LJW 52 (HKAS 97845); same location, in a subalpine forest, altitude 3000 m, 5 September 2016, KD-LJW 53 (HKAS 97846); same location, in a subalpine forest, altitude ca. 3000 m, 8 September 2016, KD-DXX 84 (HKAS 97165); Li County, Zhegushan, in a subalpine forest of Abies and Picea, altitude 3170 m, 24 June 2014, Qing Cai 1075 (HKAS 83536); same location, in a subalpine forest of Abies and Picea, altitude 3170 m, 24 June 2014, Yan-Jia Hao 1127 (HKAS 82918); Litang County, in a subalpine forest, altitude 3600 m, 13 July 2014, Jian-Wei Liu 49 (HKAS 90843); Luhuo County, in a subalpine forest, altitude 3600 m, 15 July 2014, Jian-Wei Liu 62 (HKAS 90856); same location, in a subalpine forest, altitude ca. 3600 m, 16 July 2014, Jian-Wei Liu 70 (HKAS 90864); Songpan County, in a subalpine forest, altitude 3560 m, 20 July 2014, Kuan Zhao 585 (HKAS 89514); same location, in a subalpine forest of Abies and Picea, altitude ca. 3500 m, 23 June 2014, Qing Cai 1071 (HKAS 83532). YUNNAN PROVINCE: Yulong Naxi Autonomous County, in a subalpine forest with Quercus and Abies, altitude 3500 m, 3 August 1995, Zhu L. Yang 2132 (HOLOTYPE, HKAS 29508).

Specimens of A. aff. brunneofuliginea examined: AUSTRIA: 26 September 2016, Zhu L. Yang 5914 (HKAS 96170); same location, 26 September 2016, Zhu L. Yang 5916 (HKAS 96171); same location, 26 September 2016, Zhu L. Yang 5919 (HKAS 96174).

Commentary: Amanita brunneofuliginea was described from China by Yang (1997). In our multi-locus phylogenetic data (Fig. 2), specimens of A. aff. brunneofuliginea from Europe and A. brunneofuliginea specimens from Asia split into two sister clades. These were identified as two phylogenetic species based on GCPSR analysis, but combined as a single phylogenetic species based on PTP analysis.

Collections of A. aff. brunneofuliginea have an umbonate, gray to grayish pileus ca. 5 cm in diam., striations ca. 0.2–0.3 R on the pileal margin, cream to whitish lamellae, a white to grayish stipe ca. 11–15 cm long × 0.5–1 cm diam., a saccate, easily broken volva with its outer surface dirty white with yellowish to ocheraceous spots and its inner surface dirty white, globose to subglobose basidiospores measuring [60/3/3] 11.0–13.0 × 10.0–12.0 (–12.5) μm, Q = 1.0–1.1, Qm = 1.05 ± 0.03, an absence of clamps and an association with trees of the genera Abies, Picea and Fagus. Amanita aff. brunneofuliginea can be distinguished from A. brunneofuliginea by its relatively smaller basidioma, a paler colored (grayish to gray) pileus, and globose to subglobose basidiospores. In this study, we therefore treat these taxa as two separate species: A. aff. brunneofuliginea and A. brunneofuliginea.

Further study and more collections are necessary before A. aff. brunneofuliginea can be formally described. For detailed descriptions, comparisons with similar species, line drawings and images of A. brunneofuliginea see Yang (1997, 2005, 2015).

50. Amanita brunneoprocera Thongbai & Raspé & K.D. Hyde, Mycosphere 9(3): 471 (2018).

Basidioma medium-sized. Pileus 6–8 cm diam., convex, plano-convex to applanate, dark gray (1D1–4), gray-brown (2C2–4) to gray (1C1–3, 1D1–4); volval remnants on pileus absent; margin striate (ca. 0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe ca. 13–16 cm long × 0.9–1.4 cm diam., subcylindric or slightly tapering upward, with apex slightly expanded, white (1A1) to dirty white (2B1), densely covered with minute, gray (1C1–3) to dark gray (1D1–4) fibrils; context white (1A1), hollow in center; basal bulb absent; volval remnants on stipe base saccate, 1.5–3 cm high × 1.5–2 cm wide, membranous, both surfaces white (1A1). Annulus absent. Odor indistinct.

Basidia 40–65 × 14–18 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores [120/3/2] 9.0–11.0 (–12.0) × (8.0–) 8.5–10.0 (–11.0) μm, Q = 1.00–1.14 (–1.22), Qm = 1.07 ± 0.05, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Clamps absent in all parts of basidioma.

Habit: Solitary to scattered in broad-leaved or mixed forests; basidioma occurring in summer.

Distribution: Known from southwestern China and Thailand (Thongbai et al. 2018).

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lancang County, in a mixed forest with Fagaceae and Pinaceae, altitude 1000 m, 19 August 2016, LC-LJW 10 (HKAS 97514); same location, in a subtropical forest, altitude 1370 m, 25 September 2016, LC-WPM 31 (HKAS 98435).

Commentary: Amanita brunneoprocera was described from Thailand by Thongbai et al. (2018). This species is characterized by its dark gray, gray-brown to gray pileus, a white stipe covered with gray to dark gray fibrils and globose to subglobose basidiospores (9.0–11.0 × 8.5–10.0 μm).

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. brunneoprocera is related to A. orienticrocea Zhu L. Yang, Yang-Yang Cui & Qing Cai and A. albidostipes Yang-Yang Cui et al. However, A. orienticrocea has brownish to yellow-brown pileus and a white stipe covered with indistinct squamules. For comparisons between the A. brunneoprocera and A. albidostipes see the commentary on A. albidostipes.

Morphologically, A. brunneoprocera is similar to A. ovalispora Boedijn. However, A. ovalispora has longer striations on the pileal margin (0.3–0.5 R) and narrower basidiospores (9.0–11.0 × 7.0–9.0 μm, Q = 1.13–1.38, Qm = 1.25 ± 0.08) (Boedijn 1951; Yang et al. 2001; Yang 2005, 2015).

51. Amanita changtuia Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 15b, c, 17.
Fig. 17

Microscopic features of Amanita changtuia (TYPE, HKAS 92100). a Hymenium and subhymenium; b basidiospores; c longitudinal section of interior part of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825006

Etymology: changtuia from changtui, it means long stipe in Chinese, referring to the long stipe of the basidioma representing this species.

Type: CHINA. HEILONGJIANG PROVINCE: Mudanjiang, in a temperate mixed forest with Fagaceae and Pinaceae, altitude 696 m, 1 September 2015, Qing Cai 1482 (TYPE, HKAS 92100, GenBank Acc. No.: ITS = MH508299, nrLSU = MH486442, tef1-α  = MH508724, rpb2 = MH485919).

Basidioma (Fig. 15b, c) medium-sized. Pileus 5–6.5 cm diam., applanate, umbonate and dark brown (1E5–8) at center, but becoming brown (2C3–6) to brownish (2B2–6) towards margin; volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edges white (1A1); lamellulae truncate, plentiful. Stipe 15–22.5 cm long × 1–1.5 cm diam., slender, subcylindric or slightly tapering upwards, with apex slightly expanded, white (1B1), dirty white (2B1) to brownish (1B1–3), densely covered with minute, concolorous squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 2–2.5 cm high × 1–1.5 cm wide, membranous, both surfaces white (1A1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 15–40 μm wide, composed of abundant, ellipsoid inflated cells (60–110 × 20–60 μm); filamentous hyphae abundant, 3–7 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid to fusiform inflated cells (35–90 × 15–25 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 17a) 30–50 μm thick, with 2–3 layers of subglobose, ovoid, ellipsoid to fusiform or irregularly arranged cells, 10–20 × 10–15 μm. Basidia (Fig. 17a) 50–70 × 12–18 μm, clavate, 4-spored; sterigmata 4–7 μm long; basal septa lacking clamps. Basidiospores (Fig. 17b) [120/3/2] (9.0–) 9.5–11.5 (–13.0) × 9.0–11.0 (–12.0) μm, Q = 1.00–1.14 (–1.22), Qm = 1.07 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate, inflated cells (10–30 × 10–20 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 50–90 μm thick; upper layer (15–40 μm thick) gelatinized, composed of radially arranged to interwoven, thin-walled, colorless to yellowish, filamentous hyphae 2–5 μm wide; lower layer (35–50 μm thick) composed of radially arranged, filamentous hyphae 3–6 μm wide, yellowish to yellow-brown; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 17c) composed of more or less longitudinally arranged elements: filamentous hyphae abundant, 2–11 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, globose, subglobose, ellipsoid to fusiform, 40–90 × 30–70 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer and inner surface of volval remnants on stipe base similar to structure of interior part, but with inner surface gelatinized. Stipe trama composed of longitudinally arranged, clavate terminal cells, 100–350 × 15–50 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in temperate mixed forests with Fagaceae and Pinaceae; basidioma occurring in autumn.

Distribution: Known from northeastern China.

Additional specimen examined: CHINA. HEILONGJIANG PROVINCE: Mudanjiang, in a temperate mixed forest with Fagaceae and Pinaceae, altitude 700 m, 1 September 2015, Qing Cai 1487 (HKAS 92105).

Commentary: Amanita changtuia is characterized by its slender basidioma with an umbonate pileus, globose to subglobose basidiospores (9.5–11.5 × 9.0–11.0 μm), and distribution in temperate forests.

Our ITS phylogenetic analysis (Fig. S8) indicates that A. changtuia is sister to A. constricta Thiers & Ammirati and A. cinnamomea Thongbai et al. However, A. constricta has a relatively larger and robuster basidioma with a gray to brownish gray pileus ca. 5–13 cm in diam., lamellae at first white and then becoming gray, and gray lamellar edges. Moreover, A. constricta occurs during the winter months in the USA (Thiers and Ammirati 1982; Jenkins 1986; Tulloss 1994). Amanita cinnamomea has cinnamon lamellar edges and distributed in tropical forests with trees of Fagaceae (Thongbai et al. 2018).

Amanita tenuifulva, a novel species described in this study, is easily confused with A. changtuia. However, A. tenuifulva has a non-umbonate pileus and is found in the southern regions of China. The European A. fulva is similar to A. changtuia in the color of its pileus and the similar basidiospores. However, A. fulva has a relatively shorter stipe, and a brownish, saccate volva with inflated cells dominant in the outer part (Contu 2000a, b; Massart 2000; Yang et al. 2004; Yang 2005, 2015; this study).

Amanita orientifulva also somewhat resembles A. changtuia in appearance. However, A. orientifulva has a robuster and larger basidioma, a brownish volva with abundant inflated cells in its inner part, and relatively larger basidiospores (10–14 × 9.5–13 μm) (Yang et al. 2004; Yang 2005, 2015).

52. Amanita chiui Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 15d–f, 18.
Fig. 18

Microscopic features of Amanita chiui (TYPE, HKAS 76328). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825007

Etymology: The epithet “chiui” is in honor of the late Prof. W.F. Chiu, for his early contribution to the taxonomy of Amanita in China.

Type: CHINA. SICHUAN PROVINCE: Yanyuan County, Mianyalinchang, in a mixed forest with Fagaceae and Pinaceae, altitude 2400 m, 30 July 2012, Yan-Jia Hao 670 (TYPE, HKAS 76328, GenBank Acc. No.: ITS = MH508303, nrLSU = MH486447, tef1-α  = MH508727, rpb2 = MH485930, β-tubulin  = MH485453).

Basidioma (Fig. 15d–f) medium-sized. Pileus 5–9 cm diam., convex, plano-convex to applanate, often dark gray (1F1–5), gray-brown (1E3–6), brown (1C4–8, 1D4–8) to brownish (1B2–5) over entire disk, sometimes darker at center, but paler toward margin; volval remnants on pileus mostly absent, or occasionally retained as small, white (1A1) patches; margin striate (0.25–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (3A2–3); lamellar edge white (1A1); lamellulae truncate, plentiful. Stipe 8–14 cm long × 0.5–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1B1), dirty white (2B1), brownish (1B2–5) to brown (1C4–8, 1D4–8), densely covered with dirty white (1A1) to brownish (1B2–5) squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 2.5–5.5 cm high × 1–2.5 cm wide, membranous, both surfaces white (1B1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant, ellipsoid to fusiform inflated cells (35–100 × 15–35 μm); filamentous hyphae abundant, 3–7 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid to elongate inflated cells (25–75 × 15–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 4–6 μm wide. Subhymenium (Fig. 18a) 30–50 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 15–25 × 10–20 μm. Basidia (Fig. 18a) 45–65 × 14–17 μm, clavate, 4-spored; sterigmata 4–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 18b) [120/6/4] (9.0–) 10.0–12.5 (–13.0) × (8.0–) 9.0–11.0 μm, Q = (1.01–) 1.06–1.29 (–1.33), Qm = 1.18 ± 0.07, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to fusiform or sphaeropedunculate inflated cells (10–35 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 5–10 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 90–125 μm thick; upper layer (25–60 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless, filamentous hyphae 2–6 μm wide; lower layer (50–90 μm thick) composed of radially arranged, filamentous hyphae 3–9 μm wide, colorless to yellow; vascular hyphae scarce. Volval remnants on stipe base dominantly composed of two parts intergrading to each other. Outer part of volval remnants on stipe base (Fig. 18c) composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–7 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, but becoming fairly abundant toward inner part, subglobose, ellipsoid to fusiform, 30–60 × 25–55 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Inner part of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae abundant to very abundant, 2–13 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant, subglobose, ellipsoid to fusiform, 40–80 × 15–50 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of outer part, but with more abundant, filamentous hyphae; inner surface gelatinized and similar to structure of inner part. Stipe trama composed of longitudinally arranged, clavate terminal cells, 60–400 × 25–50 μm; filamentous hyphae scattered to abundant, 3–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from northwestern and southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a forest dominated by Fagaceae, altitude 2500 m, 10 July 2008, Li-Ping Tang 382 (HKAS 54613); Lanping County, in a forest with Pinus yunnanensis, altitude 2600 m, 15 August 2010, Xue-Tai Zhu 162 (HKAS 68338). GANSU PROVINCE: Chengxian County, in a forest with Cyclobalanopsis, altitude 1260 m, 21 August 2011, Xue-Tai Zhu 521 (HKAS 73916).

Commentary: Amanita chiui is distinguished by its dark gray, brown to brownish pileus, a white to dirty white stipe often densely covered with brownish squamules, and subglobose to broadly ellipsoid basidiospores (10.0–12.5 × 9.0–11.0 μm).

In our multi-locus phylogenetic data (Fig. 2), specimens labeled as A. chiui split into two subclades (A. chiui subclade and HKAS 54613 subclade), which were recognized as a single phylogenetic species by the GCPSR analysis and as two phylogenetic species by the PTP analysis. The single collection in the HKAS 54613 clade has a gray-brown pileus with a dark center, striations on the pileal margin ca. 0.35 R, white lamellae with gray edges, a gray pileus decorated with brownish squamules, a white saccate volva on the stipe base, subglobose to broadly ellipsoid basidiospores measuring [40/1/1] 10.0–12.0 (–13.0) × 9.0–10.0 μm, Q = 1.08–1.28 (–1.33), Qm = 1.18 ± 0.05, no clamps and is in association with trees of the families Fagaceae and Pinaceae. No stable morphological differences were observed between fungi in the A. chiui clade and the single specimen of the HKAS 54613 clade, thus for the present, we treat HKAS 54613 as A. chiui.

Morphologically, A. fulva Fr. can be confused with A. chiui. However, A. fulva, described from Europe, differs from A. chiui by its glabrous stipe surface, and a saccate volva on the stipe base, with inflated cells dominant in the outer part of the volva (Contu 2000a, b; Massart 2000; Yang et al. 2004; Yang 2005, 2015; this study).

Amanita orientifulva Zhu L. Yang et al. is also similar to A. chiui, but A. orientifulva has relatively larger, globose to subglobose basidiospores (10.0.–14.0 × 9.5–13.0 μm), and occurs in subalpine forests (Yang et al. 2004; Yang 2005, 2015).

Amanita tenuifulva Yang-Yang Cui et al., a novel species described in this study, looks like A. chiui, but A. tenuifulva has a slender and smaller basidioma with a shorter stipe, globose to subglobose basidiospores (10.0–12.5 × 9.0–11.0 μm), and a saccate volva on the stipe base with very abundant inflated cells in its outer part.

53. Amanita cinctipes Corner & Bas, Persoonia 2(3): 299 (1962).

Pileus 5–7 cm in diam., sometimes umbonate, dark gray (2E1–3) to gray-brown (1E4–7) at center, but gray (1C1–3) at margin; volval remnants on pileus removable, verrucose to felted, dark gray (2E1–3) to gray (1C1–3); margin striate (0.3–0.4 R), non-appendiculate. Lamellae white (1A1) to grayish (2B1); lamellulae truncate. Stipe 6–13 cm long × 0.5–1.5 cm diam., dirty white (1B1) to grayish (2B1), covered with fibrous to pulverulent, gray (1C1–3) squamules; basal bulb absent; volval remnants on stipe base floccose in belts, gray (1C1–3) to grayish (2B1). Annulus absent.

Basidia 35–55 × 10–15 μm, clavate, 4-spored. Basidiospores [80/2/2] (8.5–) 9.0–10.5 × 8.0–9.5 (–10.0) μm, Q = 1.0–1.16 (–1.18), Qm = 1.08 ± 0.04, globose to subglobose, inamyloid. Volval remnants on pileus composed of vertically arranged, fairly abundant filamentous hyphae and very abundant inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests; basidioma occurring in summer and autumn.

Distribution: Known from southern China (Yang 2015; this study), Malaysia (Corner and Bas 1962; Lee 2017) and Singapore (Corner and Bas 1962).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude ca. 450 m, 16 May 2012, Fang Li 280 (HKAS 101388); same location, in a subtropical forest dominated by Fagaceae, altitude ca. 450 m, 23 May 2012, Fang Li 354 (HKAS 78465).

Commentary: The type of A. cinctipes was collected from Singapore, and this species has also been collected from Malaysia (Corner and Bas 1962; Lee 2017). The Chinese collections match well with the morphological and ecological characters of A. cinctipes, and, thus, Yang (2015) treated HKAS 78465 as A. cinctipes. In this study, we follow the treatment of Yang (2015), and generated DNA sequences from the Chinese materials.

Phylogenetically, our data suggest that there are no species closely related to A. cinctipes (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. cinctipes see Yang (2015).

54. Amanita cingulata J. W. Liu & Zhu L. Yang, Phytotaxa 326 (1): 47 (2017).

Pileus 4–5.5 cm in diam., plano-convex to applanate, white (1A1) to cream (3A1–2); volval remnants on pileus cracking into irregular, cushion-like or nubbly patches with maturity, white (1A1); margin striate (0.2–0.3 R), non-appendiculate. Lamellae white (1A1) to cream (3A1–2); lamellulae truncate. Stipe 6–8 cm long × 0.9–1.7 cm diam., white (5A1) to cream (5A2), covered with snakeskin-shaped, white squamules above annulus, and decorated with minute, sometimes recurved floccules to fibrils under annulus; basal bulb absent; volval remnants on stipe base saccate, but breaking into lumpy volval remnants as on pileus, white (1A1). Annulus present, thin, membranous, white (1A1), finely and radially radiate above, nearly smooth below, superior, easily detached from stipe.

Basidia 43–50 × 7–11 μm, clavate, 4-spored. Basidiospores [200/2/2] (7.5–) 9.0–11.5 (–12.0) × (5.0–) 5.5–7.0 (–8.5) μm, Q = (1.25–) 1.4–1.83 (–2), Qm = 1.58 ± 0.13, ellipsoid to elongate, rarely broadly ellipsoid, inamyloid. Volval remnants on pileus composed of ± irregularly arranged elements: filamentous hyphae fairly abundant; inflated cells very abundant to locally dominant. Volval remnants on base similar to those on pileus, but with more abundant filamentous hyphae. Annulus predominantly composed of very abundant to nearly dominant filamentous hyphae and scarce inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central and eastern China (Liu et al. 2017; this study).

Specimens examined: CHINA. HUNAN PROVINCE: Zhuzhou County, Yanling Scenic Spot, in a mixed forest dominated by Fagaceae and Theaceae, altitude 1516 m, 22 July 2012, Qing Cai 866 (HOLOTYPE, HKAS75600). JIANGXI PROVINCE: Jiujiang County, Lushan Scenic Spot, in a mixed forest dominated by Fagaceae and Pinaceae, altitude 950–994 m, 21 July 2017, Gang Wu 2185 (HKAS100640).

Commentary: Amanita cingulata is currently the only annulate species of A. sect. Vaginatae from China (Liu et al. 2017). It is characterized by its white basidioma with white, patchy volval remnants attached to its pileus, an annulate stipe, a saccate volva on the stipe base, ellipsoid to elongate basidiospores (9.0–11.5 × 5.5–7.0 μm) and the absence of clamps. For detailed descriptions, comparisons with similar species, line drawings and images of A. cingulata see Liu et al. (2017).

55. Amanita griseofolia Zhu L. Yang, Frontiers Basidiomycote Mycology: 315 (2004).

Amanita ceciliae sensu Zhu L. Yang, Bibl. Mycol. 170: 116 (1997), p.p.

Pileus 3–7 (–9) cm in diam., non- or slightly umbonate, brown-gray (1E4–7), dark gray (2E1–3) to gray (1C1–3); volval remnants on pileus removable, conical to felted, dark gray (2E1–3) to gray (1C1–3); margin striate (0.2–0.5 R), non-appendiculate. Lamellae white (1A1) to grayish (2B1); lamellulae truncate. Stipe 8–16 cm long × 0.5–1.5 cm diam., white (1A1) to dirty white (1B1), covered with fibrous to pulverulent, gray (1C1–3) squamules; basal bulb absent; volval remnants on stipe base pulverulent, irregularly arranged or in belts, dark gray (2E1–3) to gray (1C1–3).

Basidia 45–70 × 15–20 μm, clavate, 4-spored. Basidiospores [366/18/8] (9.5–) 10.0–13.5 (–16.5) × (8.5–) 9.5–13.0 (–15.0) μm, Q = 1.0–1.1 (–1.18), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid. Volval remnants on pileus composed of vertically arranged, fairly abundant filamentous hyphae and very abundant inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China (Yang 2004, 2005, 2015; this study), India (Bhatt et al. 2017) and Republic of Korea (Kim et al. 2013b).

Specimens examined: CHINA. YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a subtropical broad-leaved forest, altitude 2300 m, 6 August 2013, Zhu L. Yang 5737 (HKAS 80017); Kunming, Heilongtan, in a forest dominated by Fagaceae, altitude 1900 m, 13 August 2016, Jian-Wei Liu 545 (HKAS 98124); same location, in a forest dominated by Quercus and Pinus, altitude 2000 m, 27 June 2001, Zhu L. Yang 3081 (HOLOTYPE, HKAS 38159); same location, in a forest with Taxodiaceae, altitude 1990 m, 16 August 2008, Zhu L. Yang 5146 (HKAS 54443); Lancang County, in a mixed forest with Fagaceae and Pinaceae, altitude 1450 m, 22 August 2016, LC-Hao 64 (HKAS 96901); same location, in a forest with Fagaceae, altitude 1490 m, 23 August 2016, LC-Hao 93 (HKAS 96928); Menghai County, in a mixed forest with Fagaceae and Pinaceae, altitude 1100 m, 1 July 2014, Kuan Zhao 484 (HKAS 89147). JAPAN. HOKKAIDO: Tomakomai, in a forest of Quercus mongolica, altitude 300 m, 15 September 2011, Zhu L. Yang 5570 (HKAS 71055).

Specimen of A. ceciliae examined: FRANCE. JURA: near Noga, 2 October 1992, C. Bas 9341 (L).

Specimens of A. submembranacea examined: AUSTRIA. VORARLBERG: Kleinwalsertal, in a forest with Abies and Picea, altitude 1200 m, 29 September 2016, Zhu L. Yang 5958 (HKAS 96167); same location, altitude 1150 m, 6 September 2004, KR 8530 (MB-001175). GERMANY. FREE STATE OF BAVARIA: Oberstdorf, altitude 1120 m, 8 September 2004, KR 8584 (MB-001174).

Commentary: Amanita griseofolia was described from China by Yang (2004), and subsequently reported from India and the Republic of Korea (Kim et al. 2013b; Bhatt et al. 2017). In China, specimens of A. griseofolia have been identified as A. ceciliae (Berk. & Broome) Bas (Yang 1997). Indeed, A. griseofolia is closely related to A. ceciliae (Fig. 1b). However, the European A. ceciliae differs from A. griseofolia by its brown pileus with a yellow tinge, white lamellae that do not change to gray when dried and more filamentous hyphae in the volval remnants on the pileus (Phillips 1990; Breitenbach and Kränzlin 1995; Massart 2000; Yang 2004, 2005, 2015; this study).

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. griseofolia is also somewhat related to A. submembranacea (Bon) Gröger, but the latter has a brown, brownish to buff pileus with an olivaceous tinge, volval remnants on the pileus are absent or retained as patches mostly concentrated over the disc, a saccate volva on the stipe base and distributions in forest dominated by Abies and Picea (Tulloss 1994; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; this study).

For detailed descriptions, comparisons with similar species, line drawings and images of A. griseofolia see Yang (2004, 2005, 2015).

56. Amanita griseoumbonata Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 15g–i, 19.
Fig. 19

Microscopic features of Amanita griseoumbonata (TYPE, HKAS 92103). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base; d longitudinal section of inner part of volval remnants on stipe base. Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825008

Etymology: griseoumbonata, from griseus = gray, umbonatus = umbonate, referring to its gray distinctly umbonate pileus.

Type: CHINA. HEILONGJIANG PROVINCE: Mudanjiang, in a mixed forest with Fagaceae and Pinaceae, altitude 696 m, 1 September 2015, Qing Cai 1485 (TYPE, HKAS 92103, GenBank Acc. No.: ITS = MH508389, nrLSU = MH486578, tef1-α  = MH508847, rpb2 = MH486040).

Basidioma (Fig. 15g–i) small to medium-sized. Pileus 4–6 cm diam., plano-convex to applanate, distinctly umbonate at center, dark brown (1D3–7), gray-brown (1C3–6) to gray (2C2–5); volval remnants on pileus mostly absent, or occasionally retained as white patches; margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edge white (1A1); lamellulae truncate, plentiful. Stipe 9–15 cm long × 0.8–1.2 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), glabrous or covered with minute, concolorous fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 2–3.5 cm high × 1–2 cm wide, membranous, both surfaces white (1A1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 15–50 μm wide, composed of abundant, subglobose to ellipsoid, inflated cells (45–100 × 18–50 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform to ellipsoid inflated cells (50–70 × 15–35 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae abundant and 2–10 μm wide. Subhymenium (Fig. 19a) 35–50 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 10–30 × 10–20 μm. Basidia (Fig. 19a) 45–60 × 13–17 μm, clavate, 4-spored; sterigmata 5–8 μm long; basal septa lacking clamps. Basidiospores (Fig. 19b) [160/4/3] (9.0–) 10.0–12.0 × 9.0–11.0 (–12.0) μm, Q = 1.0–1.1 (–1.12), Qm = 1.05 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, fusiform to ellipsoid or sphaeropedunculate inflated cells (10–60 × 10–40 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 5–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 40–175 μm thick; upper layer [25–60 (–90) μm thick] slightly or non-gelatinized, composed of radially arranged, thin-walled, colorless to brownish, filamentous hyphae 2–6 μm wide; lower layer (30–90 μm thick) composed of radially arranged, filamentous hyphae 4–10 μm wide, brownish; vascular hyphae scarce. Volval remnants on stipe base (Fig. 19c, d) composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–9 μm wide, colorless to brownish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, subglobose, ellipsoid to fusiform, 15–60 × 13–50 μm, colorless to brownish, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of outer part, but with more abundant, filamentous hyphae; inner surface gelatinized and similar to structure of inner part. Stipe trama composed of longitudinally arranged, clavate terminal cells, 80–450 × 15–40 μm; filamentous hyphae scattered to abundant, 3–15 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in mixed forests dominated by Fagaceae and Pinaceae; basidioma occurring in summer to autumn.

Distribution: Known from central, northeastern and southwestern China.

Additional specimens examined: CHINA. HUBEI PROVINCE: Yichang, Shennongjia, in a forest of Abies and Betula, altitude 2300 m, 18 July 2012, Qing Cai 860 (HKAS 75594). YUNNAN PROVINCE: Lanping County, in a mixed forest, altitude 2600 m, 15 August 2010, Qi Zhao 867 (HKAS 69588).

Commentary: Amanita griseoumbonata is well characterized by its gray-brown to brown umbonate pileus, globose to subglobose basidiospores (10.0–12.0 × 9.0–11.0 μm), and a white, saccate volva with abundant to very abundant inflated cells.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. griseoumbonata is closely related to A. shennongjiana Yang-Yang Cui et al., a novel species described in this study. For comparisons between the two species see the commentary on A. shennongjiana.

Amanita ovalispora Boedijn may be confused with A. griseoumbonata, but A. ovalispora possesses a non-umbonate pileus, broadly ellipsoid to ellipsoid basidiospores (8.5–10.0 × 6.5–7.5 μm) and distributions in tropical to subtropical areas (Boedijn 1951; Yang et al. 2001; Yang 2005, 2015; see below).

The European A. vaginata var. vaginata resembles A. griseoumbonata in that they share a gray pileus. However, A. vaginata var. vaginata differs from A. griseoumbonata by its relatively robust basidioma and a non-umbonate pileus with shorter striations at the margin (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; this study).

Amanita pseudovaginata, a species described in Japan, but also collected in China, is morphologically comparable to A. griseoumbonata. However, A. pseudovaginata has a paler pileus and subglobose to broadly ellipsoid basidiospores (10.5–13 × 8–9 μm) (Hongo 1983; Yang 1997, 2005, 2015; Yang and Doi 1999).

57. Amanita hamadae Nagas. & Hongo, Trans. Mycol. Soc. Japan 25(4): 369 (1984).

Pileus 6–8 cm in diam., distinctly umbonate, dark brown (3E5–8) to brown (3D5–8) at center, becoming brown-yellow (3B4–8) to yellow (3A4–8) towards margin; volval remnants on pileus absent; margin striate (0.2–0.5 R), non-appendiculate. Lamellae white (1A1) to cream (1A2); lamellulae truncate. Stipe 13–17 cm long × 1–2 cm diam., white (1A1) to yellowish (2A3–5); basal bulb absent; volval remnants on stipe saccate, 2–4 cm high × 1.5–2 cm wide. Annulus absent.

Basidia 50–60 × 11–13 μm, clavate, 4-spored. Basidiospores [120/3/3] 10.0–12.0 (–13.0) × (7.5–) 8.0–9.0 (–9.5) μm, Q = (1.18–) 1.22–1.37 (–1.41), Qm = 1.3 ± 0.07, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Interior of volval remnants on pileus composed of vertically arranged, very abundant filamentous hyphae and scarce to fairly abundant inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China (Yang 2015; this study), and Japan (Nagasawa and Hongo 1984).

Specimens examined: CHINA. GUANDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 500 m, 5 September 2012, Fang Li 999 (HKAS 79076); same location, in a subtropical forest dominated by Fagaceae, altitude ca. 500 m, 7 September 2012, Fang Li 1036 (HKAS 79081). FUJIAN PROVINCE: Zhangping, in a subtropical forest of Pinus massoniana, altitude 280 m, 24 July 2013, Nian-Kai Zeng 1269 (HKAS 83451).

Commentary: Amanita hamadae was described from Japan (Nagasawa and Hongo 1984). Yang (2015) reported this taxon from China. In this study, we follow Yang’s (2015) concept, and provided DNA sequences generated from the Chinese collections.

The closely related species of A. hamadae were not found by our currently multi-locus phylogenetic analysis (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. hamadae see Yang (2015).

58. Amanita lignitincta Zhu L. Yang ex Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov. [Amanita lignitincta Zhu L. Yang, Bibl. Mycol. 170: 83 (1997), nom. prov., nom. inval.]

Figs. 15j–l, 20
Fig. 20

Features of Amanita lignitincta (TYPE, HKAS 29512). a Lamellar trama, subhymenium and hymenium; b Basidiospores; c Longitudinal section of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm (Yang 1997)

MycoBank: MB 825009

Etymology: lignitincta, from ligni = log, tincta = colored, referring to its wood colored pileus

Type: CHINA. YUNNAN PROVINCE: Lijiang, in a forest of Abies and Quercus, altitude 3500 m, 1 August 1995, Zhu L. Yang 2121 (TYPE, HKAS 29512, GenBank Acc. No.: nrLSU = AF024461).

Basidioma (Fig. 15j–l) medium-sized. Pileus 4–7 cm diam., applanate, gray-brown (2E1–3), brown (1C3–6) to brownish (3B2–5), sometimes with an indistinctly dark-colored ring-like zone at the proximal end of marginal striations; volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edge white (1A1); lamellulae truncate, plentiful. Stipe 7–10 cm long × 0.6–1 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to brownish (3B2–5), glabrous or covered with minute, concolorous fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 2–3.5 cm high × 1–1.5 cm wide, membranous, outer surface white (1A1), inner surface dirty white (2B1) to brownish (3B2–5). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–40 μm wide, composed of abundant fusiform, elongate to clavate inflated cells (45–70 × 15–20 μm); filamentous hyphae abundant, 3–5 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform, ellipsoid to elongate inflated cells (40–60 × 10–15 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 3–7 μm wide. Subhymenium (Fig. 20a) 25–35 μm thick, with 2–3 layers of ovoid, subglobose to ellipsoid or irregularly arranged cells, 8–15 × 6–12 μm. Basidia (Fig. 20a) 40–63 × 12–16 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 20b) [140/7/5] (9.5–) 10.0–13.0 (–13.5) × (8.5–) 9.0–12.0 (–13.0) μm, Q = (1.0–) 1.02–1.1 (–1.18), Qm = 1.06 ± 0.03, globose to subglobose, rarely broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, fusiform to ellipsoid or sphaeropedunculate, inflated cells (25–40 × 20–35 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–4 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 25–50 (–70) μm thick; upper layer (10–20 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless, filamentous hyphae 1–3 μm wide; lower layer (20–60 μm thick) composed of radially arranged, filamentous hyphae 2–5 (–15) μm wide, brown to dark brown; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 20c) composed of longitudinally arranged to interwoven elements: filamentous hyphae very abundant to dominant, 2–7 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, ovoid, subglobose, ellipsoid, elongate to clavate, 50–95 × 17–70 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base composed of very abundant to dominant filamentous hyphae, and scarce to not easily observed inflated cells; inner surface of volval remnants on stipe base gelatinized, composed of very abundant, filamentous hyphae, and scarce inflated cells. Stipe trama composed of longitudinally arranged, clavate, terminal cells, 180–320 × 28–40 μm; filamentous hyphae scattered to abundant, 2–7 (–12) μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subalpine forests with Abies, Picea, Quercus, and Pinus; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lijiang, in a forest with Pinaceae, altitude 2300 m, 13 July 2010, Jiao Qin 48 (HKAS 67734); Yulong Naxi Autonomous County, in a subalpine forest, altitude 3231 m, 12 July 2010, Qi Zhao 690 (HKAS 69411); same location, in a forest with Pinaceae, altitude 3231 m, 12 July 2010, Qi Zhao 687 (HKAS 69408); Shangri-la, in a forest of Quercus semecarpifolia, altitude 3650 m, 24 August 2009, Qing Cai 121 (HKAS 58788).

Commentary: Amanita lignitincta is characterized by its brown pileus with long striations on the margin, volval remnants on the stipe base composed of very abundant to dominant filamentous hyphae, globose to subglobose basidiospores (10.0–13.0 × 9.0–12.0 μm), and subalpine distributions.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. lignitincta is related to A. pallidocarnea (Höhn.) Boedijn and A. pallidozonata Yang-Yang Cui et al. However, A. pallidocarnea differs from A. lignitincta by its reddish to pink lamellae, a dirty pinkish stipe covered with concolorous squamules, and a tropical to subtropical distribution (Boedijn 1951; Yang 2001, 2005, 2015). Amanita pallidozonata has a relatively paler pileus and more inflated cells in the volval remnants on the stipe base.

Amanita zonata Yang-Yang Cui et al. can be confused with A. lignitincta, but the former has relatively shorter striations at the pileus margin, relatively smaller basidiospores (9–10.5 × 8.5–10 μm), and a tropical distribution under broad-leaved trees.

Amanita lignitincta is also similar to A. umbrinolutea. However, A. umbrinolutea has a darker colored pileus and brownish to gray-brown lamellae (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015; this study).

59. Amanita liquii Zhu L. Yang, M. Weiß & Oberw., Mycologia 96(3): 640 (2004).

Pileus 10–14 cm in diam., slightly umbonate, dark brown (3E6–8) to black (3F4–8); volval remnants on pileus verrucose to felted, dark gray (3E2–5) to gray-brown (4E2–4); margin striate (0.1–0.3 R), non-appendiculate. Lamellae white (1A1) when young, grayish (2B1) when mature, dark gray (2D1–3) to dark brown (2E1–2) when dried; lamellar edges black (3F4–8) to dark brown (2E1–2); lamellulae truncate. Stipe 13–17 cm long × 1.5–3 cm diam., brownish (3D2–4), covered with snakeskin-shaped, dark gray (3E2–5) to black (3F4–8) squamules; basal bulb absent; volval remnants on stipe verrucose to conical, arranged in incomplete rings, gray (4D1–2), dark gray (3E2–5) to brown gray (4E2–4).

Basidia 55–95 × 16–25 μm, clavate, 4-spored. Basidiospores [120/6/5] (11.0–) 11.5–15.0 (–24.0) × (9.5–) 11.0–14.5 (–20.0) μm, Q = 1.0–1.09 (–1.21), Qm = 1.05 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Verrucae on pileus composed of subvertically arranged filamentous hyphae and inflated cells. Felts on pileus composed of irregularly arranged filamentous hyphae and inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in alpine to subalpine, pine forests; basidioma occurring in summer and autumn.

Distribution: Known from eastern and southwestern China (Yang et al. 2004; Yang 2005, 2015; this study) and India (Bhatt et al. 2003; Kaur et al. 2011).

Specimens examined: CHINA. YUNNAN PROVINCE: Lijiang, Yulongxueshan, in a subalpine forest of Abies, altitude 3900 m, 14 August 2000, Zhu L. Yang 2916 (HOLOTYPE, HKAS 36611); Shangri-la, in an alpine to subalpine forest of Abies, altitude 3750 m, 25 August 2009, Qing Cai 135 (HKAS 58800); same location, in an alpine to subalpine forest of Abies, altitude ca. 3500 m, 9 July 2014, Bang Feng 1531 (HKAS 93915); Yulong Naxi Autonomous County, Laojun Mountain, in an alpine to subalpine forest, altitude 3700 m, 3 September 2009, Qing Cai 220 (HKAS 58885). TAIWAN PROVINCE: Nantou County, Hehuanshan, in a forest of Abies, altitude 3000 m, 15 September 2012, Bang Feng 1264 (HKAS 82406).

Commentary: Amanita liquii was described from China by Yang et al. (2004), and subsequently reported from India (Bhatt et al. 2003; Kaur et al. 2011). Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. liquii is related to A. griseofolia, A. submembranacea and A. ceciliae. For discussions between A. liquii and A. griseofolia see the commentary on A. griseofolia. Amanita submembranacea has a brown, brownish to buff pileus with an olivaceous tinge, volval remnants on the pileus absent or retained as patches mostly concentrated over disc and a saccate volva on the stipe base (Tulloss 1994; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; this study). The European A. ceciliae differs from A. liquii in its brownish yellow to brownish pileus, white lamellae that do not change to gray when dried, a white stipe, and volval remnants on stipe base broken into fragments or forming a cup-like volva (Phillips 1990; Breitenbach and Kränzlin 1995; Massart 2000; Yang 2004, 2005, 2015). For detailed descriptions, comparisons with similar species, line drawings and images of A. liquii see Yang et al. (2004) and Yang (2005, 2015).

60. Amanita neocinctipes Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 15m–o, 21.
Fig. 21

Microscopic features of Amanita neocinctipes (TYPE, HKAS 79627). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825010

Etymology: neocinctipes, from neo- = new, cinctipes =the species of Amanita cinctipes, referring its similarity with A. cinctipes.

Type: CHINA. GUANGDONG PROVINCE: Fengkai County, in a subtropical forest dominated by Fagaceae (Castanopsis and Lithocarpus), altitude 250 m, 3 June 2013, Qing Cai 948 (TYPE, HKAS 79627, GenBank Acc. No.: nrLSU = MH486653, tef1-α  = MH508910, rpb2 = MH486103).

Basidioma (Fig. 15m–o) medium-sized. Pileus 4–7 cm diam., applanate, gray-brown (1D2–5) to dark gray (1E1–3) over entire disk, or often dark gray (1E1–3) at center, and becoming brown-gray (1C2–4), gray (2D2–4) to grayish (2C2–3, 2B2–3) towards margin; volval remnants on pileus pyramidal, floccose to patchy, dark gray (1E1–3) to gray (2D2–4); margin striate (0.3–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe 8–11 cm long × 0.3–1 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, upper half dirty white (2B1) to grayish (2C2–3, 2B2–3) and covered with concolorous, pulverulent squamules, lower half grayish (2C2–3, 2B2–3) to gray (2D2–4) and decorated with fibrous squamules; context white (1A1) to dirty white (2B1), hollow in center; basal bulb absent; volval remnants on stipe base floccose to felted, gray (2D2–4) to dark gray (1E1–3), arranged irregularly or in incomplete belts or rings on stipe base. Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant ellipsoid to fusiform inflated cells (45–65 × 25–40 μm); filamentous hyphae abundant, 2–7 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform, ellipsoid to elongate inflated cells (35–65 × 12–35 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 21a) 30–50 μm thick, with 2–3 layers of subglobose, fusiform, ellipsoid or irregularly arranged cells, 15–25 × 10–20 μm. Basidia (Fig. 21a) 40–60 × 11–15 μm, clavate, 4-spored; sterigmata 4–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 21b) [80/2/2] 8.0–10.5 (–11.0) × 7.0–9.0 μm, Q = 1.09–1.29 (–1.39), Qm = 1.19 ± 0.07, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of ellipsoid to subglobose, fusiform to ellipsoid or sphaeropedunculate inflated cells (10–40 × 10–30 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 60–100 μm thick; upper layer (15–40 μm thick) gelatinized, composed of interwoven to radially arranged, thin-walled, colorless to yellowish, filamentous hyphae 2–6 μm wide; lower layer (35–65 μm thick) composed of radially arranged, filamentous hyphae 3–8 μm wide, yellow brown; vascular hyphae scarce. Volval remnants on pileus (Fig. 21c) composed of vertically to somewhat irregularly arranged elements: filamentous hyphae scarce, but becoming fairly abundant toward pileipellis, 2–10 μm wide, yellow-brown to brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to nearly dominant, subglobose, subfusiform to ellipsoid, 15–80 × 10–50 μm, yellow-brown to brown, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base similar to structure of volval remnants on pileus. Stipe trama composed of longitudinally arranged, clavate terminal cells, 90–300 × 15–35 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in a subtropical forests dominated by Fagaceae; basidioma occurring in summer.

Distribution: Known from southern China.

Additional specimen examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 230 m, 22 May 2012, Fang Li 336 [HKAS 78463, as Amanita sp. 10 in Yang (2015)].

Commentary: Amanita neocinctipes is well circumscribed by its gray to gray-brown pileus densely covered with pyramids or patches, floccose volval remnants on the stipe base irregularly or arranged in incomplete belts or rings, and subglobose to broadly ellipsoid basidiospores (8.0–10.5 × 7.0–9.0 μm). Furthermore, it is associated with trees of the family Fagaceae.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. neocinctipes is related to A. lippiae Wartchow & Tulloss. However, A. lippiae has a pale beige pileus covered with numerous scattered white patches, no annulus or an apparent small subbasal pendant annulus, mostly collar-like volval remnants on the stipe base, ellipsoid to elongate basidiospores (10.0–21.0 × 7.0–11.0 μm), and is found in sandy soil under Lippia trees (Wartchow et al. 2009).

To date, four species of A. sect. Vaginatae with a non-saccate volva were described from China, including A. cinctipes, A. griseofolia, A. liquii and A. neocinctipes. The former three species can be separated from A. neocinctipes by molecular phylogenetic, morphological and ecological evidence. Amanita cinctipes differs from A. neocinctipes in its globose to subglobose basidiospores (8.7–11.5 × 7.9–10.1 μm) (Corner and Bas 1962; this study). Amanita griseofolia can be distinguished from A. neocinctipes by its globose to subglobose basidiospores (10.0–13.5 × 9.5–13.0 μm), and its distribution in mixed forests (Yang 2004, 2005, 2015). Amanita liquii is different from A. neocinctipes by its relatively larger and darker basidioma, larger basidiospores (11.5–15.0 × 11.0–14.5 μm) and its subalpine distribution (Yang et al. 2004; Yang 2005, 2015).

61. Amanita nivalis Grev., Scott. crypt. fl. (Edinburgh) 1(4): pl. 18 (1825).

Pileus 3–6.5 cm in diam., slightly umbonate, grayish (1C1–3), grayish brown (2C2–4) to dirty white (2B1); volval remnants on pileus absent; margin striate (0.1–0.4 R), non-appendiculate. Lamellae white (1A1) to grayish (1C1–3); lamellar edges brown (1D6–8) to gray (2D1–2); lamellulae truncate. Stipe 4–8 cm long × 0.5–1.2 (–1.5) cm diam., dirty white (2B1), covered with minute, grayish (1C1–3), gray (2D1–2) to brownish (1C3–5) squamules; basal bulb absent; volval remnants on stipe saccate, 2–4 cm high × 1–2.5 cm wide. Annulus absent.

Basidia 50–78 × 16–20 μm, clavate, 4-spored. Basidiospores [190/13/12] (9.0–) 9.5–12.0 (–14.0) × (8.5–) 9.0–10.5 (–13.5) μm, Q = 1.0–1.08 (–1.12), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subalpine areas near the snow-line and with Polygonum, Potentilla, Rhododendron, Cyperus, Juniperus; basidioma occurring in summer and autumn.

Distribution: Known from northern and southwestern China (Yang 2005, 2015; this study), and Europe (Gilbert 1940, 1941a; Watling 1985; Campo and Bizio 2000; Contu 2000b; Galli 2001; Neville and Poumarat 2004; Tulloss et al. 2015).

Specimens examined: CHINA. INNER MONGOLIA: Aershan, altitude unknown, 8 August 1991, Yu-Chen Zong et al. 14 (HMAS 59848 and 61226). YUNNAN PROVINCE: Shangri-La, on grassland near the snow-line with Polygonum and Potentilla, sometimes with Rhododendron, Cyperus and Juniperus, altitude 4300 m, 25 July 1998, Zhu L. Yang 2440 (HKAS 32534); same location, in grassland near the snow-line with Polygonum and Potentilla, sometimes with Rhododendron, Cyperus and Juniperus, altitude 4300 m, 25 July 1998, Zhu L. Yang 2441 (HKAS 32533); same location, in grassland near the snow-line with Polygonum and Potentilla, sometimes with Rhododendron, Cyperus and Juniperus, altitude 4300 m, 25 July 1998, Zhu L. Yang 2442 (HKAS 32528); same location, in grassland near the snow-line with Polygonum and Potentilla, sometimes with Rhododendron, Cyperus and Juniperus, altitude 4300 m, 25 July 1998, Zhu L. Yang 2445 (HKAS 32469); same location, in grassland near the snow-line with Polygonum and Potentilla, sometimes with Rhododendron, Cyperus and Juniperus, altitude 4300 m, 25 July 1998, Zhu L. Yang 2447 (HKAS 32454); same location, altitude 4300 m, 25 July 1998, Zhu L. Yang 2448 (HKAS 32453); same location, in areas near the snow-line, altitude 4300 m, 25 July 1998, Zhu L. Yang 2449 (HKAS 32452). TIBET AUTONOMOUS REGION: Bomê County, in forest near the snow-line, altitude 3000 m, 14 September 1982, Xiao-Lan Mao 455 (HMAS 53380). U.K. SCOTLAND: altitude ca. 2000 m, 20 August 1992, C. Bas 9307 (L); Grampian Reg., Cairnwell, 8 September 1984, R. Watling 17489 (ISO-EPITYPE, L).

Commentary: Amanita nivalis was described from the U. K. and then reported from various other European countries (Gilbert 1940, 1941a; Watling 1985; Campo and Bizio 2000; Contu 2000b; Galli 2001; Neville and Poumarat 2004). Watling (1985) proposed the collection R. Watling 17489 (E) as a neotype of A. nivalis. Tulloss et al. (2015) proposed Watling’s neotype [R. Watling 17489 (E)] as an epitype, and suggested three iso-epitypes deposited in L (0053705), NY (01937400), and RET (042-5, fragments). The nrLSU sequence of the iso-epitype deposited in L was provided by Weiß et al. (1998).

We have examined the iso-epitype deposited in L, and the basidiospores were [20/1/1] (9.5–) 10.0–11.5 × (8.5–) 9.0–11.0 μm, Q = (1.0–) 1.02–1.14 (–1.16), Qm = 1.09 ± 0.04, globose to subglobose. Yang (2005, 2015) reported A. nivalis from China. Unfortunately, we were unable to generate DNA sequences from the Chinese specimens due to their poor condition. New material from subalpine areas near snow-line in southwestern China, from which we are able to generate DNA sequences, will shed new light on the phylogeny and taxonomy of this taxon.

62. Amanita olivaceofusca Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22a, b, 23.
Fig. 22

Fresh basidiomata of novel species in Amanita sect. Vaginatae. ab A. olivaceofusca (a HKAS 87076, b TYPE, HKAS 97581); c–d A. orienticrocea (c HKAS 80029, d TYPE, HKAS 90455); e–g A. pallidozonata (e HKAS 80934, fg TYPE, HKAS 57718); h–i A. retenta (h TYPE, HKAS 70020, i HKAS 69787); j–k A. shennongjiana (TYPE, HKAS 75553); l–m A. tenuifulva (TYPE, HKAS 87120); n–o A. zonata (n TYPE, HKAS 97240, o HKAS 97706). Bars: 2 cm

Fig. 23

Microscopic features of Amanita olivaceofusca. a Hymenium and subhymenium (TYPE, HKAS 97581); b basidiospores (TYPE, HKAS 97581); c longitudinal section of outer part of volval remnants on stipe base, left side indicates outer surface of volval remnants on stipe base (HKAS 97766). Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825025

Etymology: olivaceofusca, from olivaceus = olive-green, and fuscus = dark brown, referring to its brown to dark brown pileus with olive tinge.

Type: CHINA. YUNNAN PROVINCE: Lancang County, in a broad-leaved forest with Fagaceae, altitude 1300 m, 22 August 2016, LC-LJW 77 (TYPE, HKAS 97581, GenBank Acc. No.: ITS = MH508457, nrLSU = MH486691, rpb2 = MH486127).

Basidioma (Fig. 22a, b) small. Pileus 2–4.5 cm diam., plano-convex to applanate, dark brown (3E4–8) to gray-brown (2D4–8) at center, becoming yellow-brown (3C3–7) brown (3B3–6, 1B3–5) towards margin, with olivaceous (30A5–8) tinge; volval remnants on pileus absent; margin striate (0.2–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellulae truncate, plentiful. Stipe 6–14 cm long × 0.3–0.6 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with minute, concolorous, fibrils or floccules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 1.5–2.5 cm high × 0.8–1.5 cm wide, membranous, both surfaces white (1A1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–50 (–75) μm wide, composed of abundant subglobose to ellipsoid inflated cells (25–80 × 15–30 μm); filamentous hyphae abundant, 2–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid to fusiform inflated cells (20–55 × 10–20 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 23a) 40–70 μm thick, with 2–3 layers of subglobose, ellipsoid to fusiform or irregularly arranged cells, 10–30 × 8–20 μm. Basidia (Fig. 23a) 35–60 × 10–17 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 23b) [160/5/5] (9.5–) 10.5–13.0 (–15.0) × (8.0–) 8.5–10.0 (–11.5) μm, Q = (1.02–) 1.05–1.45 (–1.5), Qm = 1.25 ± 0.09, broadly ellipsoid to ellipsoid, sometimes subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose or sphaeropedunculate inflated cells (15–30 × 10–20 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 40–250 μm thick; upper layer (25–150 μm thick) gelatinized, composed of interwoven to radially arranged, thin-walled, colorless to brownish, filamentous hyphae 2–4 μm wide; lower layer (15–100 μm thick) composed of radially arranged, filamentous hyphae 3–7 μm wide, brownish to brown; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 23c) composed of more or less longitudinally arranged elements: filamentous hyphae abundant to very abundant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, subglobose, ellipsoid to fusiform, 40–110 × 25–80 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base (Fig. 23c) composed of very abundant to dominant, filamentous hyphae, and scarce, inflated cells; inner surface of volval remnants on stipe base gelatinized, similar to structure of interior part. Stipe trama composed of longitudinally arranged, clavate, terminal cells, 60–400 × 20–50 μm; filamentous hyphae scattered to abundant, 2–12 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China.

Additional specimens examined: CHINA. FUJIAN PROVINCE: Sanming, in a subtropical forest with Castanopsis, altitude 250 m, 8 July 2013, Yan-Jia Hao 963 (HKAS 80243). YUNNAN PROVINCE: Lancang County, in a forest dominated by Fagaceae, altitude 1300 m, 25 September 2016, LC-WPM 33 (HKAS 98437); same location, in a forest dominated by Fagaceae, altitude 1370 m, 29 September 2016, LC-LJW 262 (HKAS 97766); Puer, in a broad-leaved forest with Fagaceae, altitude 1326 m, 11 July 2014, Xiao-Bin Liu 430 (HKAS 87076).

Commentary: Amanita olivaceofusca is characterized by its small, slender basidioma, a gray-brown to brown pileus with an olivaceous tinge, broadly ellipsoid to ellipsoid basidiospores (10.5–13.0 × 8.5–10.0 μm), and associations with broad-leaved trees in tropical to subtropical forests.

Phylogenetically, A. olivaceofusca clustered as a distinct clade within A. sect. Vaginatae, but our study did not suggest any species closely related to it (Fig. 1b). Morphologically, A. angustilamellata can be easily confused with A. olivaceofusca because of its slender basidioma and tropical to subtropical distribution, but A. angustilamellata has a relatively larger basidioma, a darker colored pileus and globose to subglobose basidiospores (9.5–11 × 9.5–10.5 µm) (Boedijn 1951; Corner and Bas 1962; Pegler 1986; Yang 2005; Lee 2017).

The slender, brown basidioma of A. tenuifulva also calls to mind A. olivaceofusca. However, A. tenuifulva can be easily distinguished from A. olivaceofusca by its globose to subglobose basidiospores (10.0–12.0 × 10.0–11.5 μm) and abundant inflated cells in the outer part of the volva.

Amanita albidostipes and A. fulva are similar to A. olivaceofusca in appearance. However, the former two species possess a larger basidioma, shorter striations on the pileal margin, and globose to subglobose basidiospores [9.0–11.0 × 8.0–10.0 μm for A. albidostipes; (9.0–) 10.0–12.0 (–17.5) × (8.0–) 9.0–11.0 (–14.0) μm for A. fulva] (Contu 2000a, b; Massart 2000; Galli 2001; Yang et al. 2004; Yang 2005, 2015; this study). In addition, A. albidostipes, a novel species described in this study, has scarce inflated cells in the volval remnants on the stipe base, and is found in association with Pinus. The European species, A. fulva, has a saccate volva with abundant inflated cells in its outer part, and it is found in temperate forests (Contu 2000a, b; Massart 2000; Yang et al. 2004; Yang 2005, 2015).

63. Amanita orienticrocea Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 22c, d, 24.
Fig. 24

Microscopic features of Amanita orienticrocea (TYPE, HKAS 90455). a Hymenium and subhymenium; b basidiospores. Bars = 10 μm

MycoBank: MB 825026

Etymology: orienticrocea, from orientalis = eastern, crocea = Amanita crocea, is referring to its similarity with A. crocea and its type locality from China.

Type: CHINA. YUNNAN PROVINCE: Jingdong County, in a forest of Pinus kesiya var. langbianensis, altitude 1300 m, 30 July 2015, Qi Zhao 2581 (TYPE, HKAS 90455, GenBank Acc. No.: ITS = MH508465, nrLSU = MH486701, tef1-α  = MH508942, rpb2 = MH486133, β-tubulin  = MH485645).

Basidioma (Fig. 22c, d) medium-sized to large. Pileus 5–11 cm diam., convex to plano-convex, often umbonate at center, brownish (2B2–5), yellow-brown (4C5–8) to yellow-gray (2D5–8) at center, dirty yellow (3A2–4), cream (1A2) to brownish (2B2–5) towards margin; volval remnants on pileus absent; margin striate (0.2–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1), but with gray tinge when young; lamellar edges white (1A1); lamellulae truncate, plentiful. Stipe 7–15 cm long × 0.7–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), often covered with indistinctly, white (1A1), dirty white (1A1) to brownish (2B2–5) squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 3–5 cm high × 1.5–3 cm wide, 2–3 mm thick, membranous, outer surface white, inner surface grayish. Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant, subglobose, ellipsoid to clavate inflated cells (30–90 × 25–50 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform to ellipsoid inflated cells (25–65 × 10–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 24a) 25–40 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 8–40 × 5–30 μm. Basidia (Fig. 24a) 45–60 × 11–14 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 24b) [100/8/8] 9.0–11.0 (–13.0) × (8.0–) 9.0–11.0 (–12.0) μm, Q = 1.0–1.11 (–1.17), Qm = 1.05 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (20–40 × 15–30 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 60–125 μm thick; upper layer (30–75 μm thick) gelatinized, composed of interwoven to radially arranged, thin-walled, colorless to brownish, filamentous hyphae 2–6 μm wide; lower layer (30–70 μm thick) composed of radially arranged, filamentous hyphae 3–12 μm wide, colorless to brownish; vascular hyphae scarce. Interior of volval remnants on the stipe base composed of irregularly to longitudinally arranged elements: filamentous hyphae abundant to very abundant, 2–12 (–15) μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to fairly abundant, subglobose, ellipsoid to fusiform, 25–90 × 20–60 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Stipe trama composed of longitudinally arranged, clavate terminal cells, 65–350 × 15–30 μm; filamentous hyphae scattered to abundant, 2–12 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Specimens examined: CHINA. YUNNAN PROVINCE: Baoshan, in a forest of Pinus yunnanensis, altitude 1680 m, 12 August 2015, Pan-Meng Wang 367 (HKAS 93327); Changning County, in a forest of Pinus yunnanensis, altitude 1784 m, 24 July 2009, Li-Ping Tang 913 (HKAS 56870); Cheng County, in a forest of Cyclobalanopsis, altitude 1260 m, 21 August 2011, Xue-Tai Zhu 520 (HKAS 73915); Gongshan County, in a pine forest, altitude 2000 m, 29 July 2011, Yan-Jia Hao 279 (HKAS 71388); Lushui County, in a forest dominated by Pinus yunnanensis, altitude 1900 m, 7 August 2011, Xue-Tai Zhu 427 (HKAS 73823); same location, in a forest dominated by Pinus yunnanensis, altitude 1900 m, 7 August 2011, Yan-Jia Hao 393 (HKAS 71502); Kunming, Yeyahu, in a forest of Keteleeria evelyniana, altitude 2000 m, 1 September 2003, Zhu L. Yang 5751 [HKAS 80029, as A. cf. crocea in Yang (2015)].

Commentary: Amanita orienticrocea is characterized by its yellow to yellow-brown pileus with grayish lamellae, a dirty white stipe covered with indistinctly, dirty white to brownish squamules, globose to subglobose basidiospores (9.0–11.0 × 9.0–11.0 μm) and the absence of clamps.

In our ITS phylogenetic analysis (Fig. S8), A. orienticrocea was closely related to A. crocea (Quél.) Singer. Indeed, they are similar morphologically in that they share a yellow pileus. However, the European A. crocea differs from A. orienticrocea by its obviously saffron orange pileus, white lamellae lacking a gray tinge and a stipe decorated with distinctly orange squamules (Gilbert 1940, 1941a; Tulloss 1997, 2000a; Contu 2000a; Massart 2000; Galli 2001).

In China, A. orienticrocea is morphologically similar to A. orientifulva Zhu L. Yang, M. Weiß & Oberw. and A. albidostipes Yang-Yang Cui, Qing Cai & Zhu L. Yang. However, A. orientifulva has a dark brown to red-brown pileus, brown to brownish lamellar edges, a dirty white stipe covered with red-brown to gray-brown squamules, brown patches on the outer surface of the volva, relatively larger basidiospores (10–14 × 9.5–13 μm), and is found in subalpine forests (Yang et al. 2004; Yang 2005, 2015). Amanita albidostipes has a non-umbonate pileus, white lamellae without gray tinge when young, and the outer surface of the volva often has a brown tinge.

In addition, A. subnudipes (Romagn.) Tulloss and A. romagnesiana Tulloss are also similar to A. orienticrocea. However, A. subnudipes has an orange pileus and relatively narrower basidiospores (8.9–12.2 × 7.5–10.5 μm, Q = 1.12–1.16) (Tulloss 2000a, b). Amanita romagnesiana has a non-umbonate, bright orange-tawny pileus, relatively shorter striations on the pileal margin (≤ 0.2 R), pale cream lamellae, and a pale brownish orange stipe decorated with pale orange floccules (Tulloss 1994, 2000a, b).

64. Amanita orientifulva Zhu L. Yang, M. Weiß & Oberw., Mycologia 96(3): 643 (2004).

Pileus 5–15 cm in diam., umbonate, dark brown (4F6–8), red-brown (5E6–8) to brown (4E6–8) at center, yellow-brown (5D3–5) to yellow (4B3–5) towards margin; volval remnants on pileus absent; margin striate (0.2–0.4 R), non-appendiculate. Lamellae white (1A1) to cream (1A2); lamellar edges brown (1D6–8) to brownish (2C2–5); lamellulae truncate. Stipe 8–15 (–20) cm long × 0.5–3 cm diam., dirty white (2B1) to brownish (2C2–5), covered with red-brown (5E6–8) to gray-brown (4D1–3) squamules; basal bulb absent; volval remnants on stipe saccate, 4–6 cm high × 1.5–5 cm wide, outer surface white (1A1) with brown (1D6–8) patches, inner surface brownish (2C2–5). Annulus absent.

Basidia 4-spored; basal septa lacking clamps. Basidiospores [250/12/6] (8.5–) 10.0–14.0 (–17.0) × (8.0–) 9.5–13.0 (–16.5) μm, Q = 1.0–1.12 (–1.23), Qm = 1.06 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Volval remnants on stipe base dominantly composed of two layers intergrading to each other. Outer layer composed of very abundant to dominant filamentous hyphae and scarce to fairly abundant inflated cells. Inner layer composed of abundant filamentous hyphae and very abundant inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subalpine forest; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang et al. 2004; Yang 2005, 2015; this study), India (Kaur et al. 2011; Bhatt et al. 2017) and Republic of Korea (Kim et al. 2013b; Lee et al. 2016).

Specimens examined: CHINA. SICHUAN PROVINCE: Batang County, in subalpine forest, altitude 3500 m, 13 September 2016, Jian-Wei Liu 572 (HKAS 101433). YUNNAN PROVINCE: Kangding County, in a subalpine forest, altitude 3000 m, 7 September 2016, KD-LJW 98 (HKAS 97891); same location, in a subalpine forest, altitude 3500 m, 11 September 2016, KD-LJW 178 (HKAS 97971); Shangri-la, Bitahai, in a subalpine forest, altitude 3600 m, 20 August 2014, Qi Zhao 2171 (HKAS 87937); Shangri-la, Dongwang, in a subalpine forest with Quercus, Abies and Rhododendron, altitude 3800 m, 26 July 1998, Zhu L. Yang 2461 (HOLOTYPE, HKAS 32522); Shangri-la, Xiaozhongdian, in a forest with Quercus, altitude 3460 m, 22 August 2014, Qi Zhao 2219 (HKAS 87985).

Commentary: Amanita orientifulva was described from China by Yang et al. (2004), and subsequently reported from India and the Republic of Korea (Kaur et al. 2011; Kim et al. 2013b; Lee et al. 2016; Bhatt et al. 2017). In our multi-locus phylogenetic analysis (Fig. 1b), A. orientifulva was closely related to A. fulva and A. tenuifulva. Comparisons between A. orientifulva and A. fulva are discussed in Yang et al. (2004) and Yang (2005, 2015). The differences between A. orientifulva and A. tenuifulva are provided in the commentary on A. tenuifulva.

65. Amanita ovalispora Boedijn, Sydowia 5(3-6): 320 (1951).

Pileus 4–7 cm in diam., non- or slightly umbonate, dark gray (2E1–2) to gray (2D1–2); volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 6–10 cm long × 0.5–1.5 cm diam., dirty white (2B1) to grayish (1C1–2); basal bulb absent; volval remnants on stipe saccate, 2–4 cm high × 1.2–2.5 cm wide. Annulus absent.

Basidia 4-spored; basal septa lacking clamps. Basidiospores [162/7/7] (8.0–) 9.0–11.0 (–12.0) × (5.5–) 7.0–9.0 (–9.5) μm, Q = (1.05–) 1.13–1.38 (–1.51), Qm = 1.25 ± 0.08, broadly ellipsoid to ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical broad-leaved or mixed forest with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China (Yang 1997, 2005, 2015; this study), and Indonesia (Boedijn 1951).

Specimens examined: CHINA. FUJIAN PROVINCE: Zhangping, in a subtropical forest dominated by Fagaceae, altitude 350 m, 7 August 2013, Nian-Kai Zeng 1394 (HKAS 101406). GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 650 m, 20 July 2012, Fang Li 697 (HKAS 101394); same location, in a forest with Castanopsis and Lithocarpus, altitude 550 m, 3 June 2013, Qing Cai 946 (HKAS 79625). YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a subtropical broad-leaved forest, altitude 2400 m, 21 July 2006, Zhu L. Yang 4698 (HKAS 50495); Lanping County, in a forest with Pinus yunnanensis, altitude 2600 m, 15 August 2010, Xue-Tai Zhu 173 (HKAS 68349); Nanhua County, 10 August 2016, Zhu L. Yang 5900 (HKAS 96114). INDONESIA. JAVA: Bogor, Botanical Garden, March 1925, van Overeem s.n. (HOLOTYPE, BO, in liquid).

Commentary: Amanita ovalispora was described from Indonesia (Boedijn 1951). Yang (1997) examined the holotype, and found that the basidiospores were [40/2/1] (8.0–) 8.5–10.0 (–10.5) × (6.0–) 6.5–7.5 (–8.0) μm, Q = (1.19–) 1.25–1.42 (–1.47), Qm = 1.32 ± 0.06. To date, no sequences of A. ovalispora from its type locality are available to delimitate this species accurately. Consequently, we conservatively treat the Chinese collections, which match well with the characters of the protologue and the holotype of A. ovalispora, as this species for the time being.

Amanita ovalispora harbors a species complex. In our multi-locus phylogenetic data (Fig. 2), specimens labeled as A. ovalispora split into three subclades, which were separated into three phylogenetic species based on PTP analysis. In our ITS phylogenetic analysis (Fig. S8), A. subovalispora, a species described from Thailand (Thongbai et al. 2018), also clustered within the A. ovalispora complex. Further studies are needed to classify the A. ovalispora complex accurately. For detailed descriptions, comparisons with similar species, line drawings and images of A. ovalispora see Yang (1997, 2005, 2015).

66. Amanita pallidocarnea (Höhn.) Boedijn, Sydowia 5(3-6): 319 (1951).

Basionym: Amanitopsis vaginata var. pallidocarnea Höhn., Sber. Akad. Wiss. Wien, Math.-naturw. Kl., Abt. 1 123: 74 (1914).

Pileus 4–8 cm in diam., dark gray (2E1–2) to dark brown (1E6–8), but gray (3E1–2) at margin; volval remnants on pileus absent; margin striate (0.3–0.4 R), non-appendiculate. Lamellae red (9A4–7); lamellulae truncate. Stipe 6–13 cm long × 0.5–1.5 cm diam., dirty pinkish (9A2–4), covered with minute, concolorous squamules; basal bulb absent; volval remnants on stipe saccate, 2–4 cm high × 1.5–2.5 cm wide. Annulus absent.

Basidia 40–70 (–80) × 12–18 μm, clavate, 4-spored. Basidiospores [100/5/5] (8.5–) 9.0–12.0 (–14.5) × 8.0–11.0 (–14.0) μm, Q = 1.0–1.16 (–1.25), Qm = 1.08 ± 0.05, globose to subglobose, inamyloid. Volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae abundant to very abundant; inflated cells fairly abundant to abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forest with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China (Yang 2001, 2005, 2015; this study), and Indonesia (Boedijn 1951).

Specimens examined: CHINA. HAINAN PROVINCE: Ledong County, Jianfengling, in a tropical forest, altitude 800 m, 20 August 1999, Ming-Sheng Yuan 4373 (HKAS 34571). YUNNAN PROVINCE: Lancang County, in a forest dominated by Fagaceae, altitude 1250 m, 25 September 2016, LC-LJW 174 (HKAS 97678); same location, in a forest dominated by Fagaceae, altitude ca. 1200 m, 26 September 2016, LC-LJW 185 (HKAS 97689); same location, in a forest dominated by Fagaceae, altitude ca. 1200 m, 25 September 2016, LC-WPM 18 (HKAS 98422); Xishuangbanna, in a mixed forest with Fagaceae and Pinaceae, altitude 1200 m, 9 July 2014, Li-Hong Han 408 (HKAS 84704).

Commentary: Amanita pallidocarnea was described from Indonesia by Boedijn (1951), and also reported from southern and southwestern China in subtropical forests with broad-leaved trees (Yang 2001, 2005, 2015).

Phylogenetically, A. pallidocarnea was closely related to A. pallidozonata (Fig. 1b), and the discussion on the differences between them is given under the commentary on A. pallidozonata. For detailed descriptions, comparisons with similar species, line drawings and images of A. pallidocarnea see Yang (2001, 2005, 2015).

67. Amanita pallidozonata Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22e–g, 25.
Fig. 25

Microscopic features of Amanita pallidozonata (TYPE, HKAS 57718). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base; d longitudinal section of inner part of volval remnants on stipe base. Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825027

Etymology: pallidozonata, from pallidus = pale, zonatus = zoned, referring to its brown pileus with a distinctly pale colored ring-like zone at proximal end of marginal striations.

Type: CHINA. YUNNAN PROVINCE: Jianchuan County, in a mixed forest with Fagaceae and Pinaceae, altitude 2700 m, 29 August 2009, Gang Wu 186 (TYPE, HKAS 57718, GenBank Acc. No.: nrLSU = MH486740, tef1-α  = MH508973).

Basidioma (Fig. 22e–g) small to medium-sized. Pileus 3.5–7 cm diam., plano-convex to applanate, gray-brown (2E4–6) to brown (3C4–6) at center and margin, but forming a distinctly pale colored [brown (3C4–6) to brownish (1B3–5)] ring-like zone at proximal end of marginal striations; volval remnants on pileus mostly absent; margin striate (0.3–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edges white (1A1) to cream (1A2); lamellulae truncate, plentiful. Stipe 7–16 cm long × 0.6–1 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to slightly brownish (1B3–5), covered with minute, concolorous fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 1.5–3 cm high × 0.7–1.5 cm wide, membranous, outer surface white, often with brown (3C4–6) tinge, inner surface white (1A1) to dirty white (2B1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant, subglobose to ellipsoid inflated cells (45–100 × 20–70 μm); filamentous hyphae abundant, 3–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, subglobose to ellipsoid inflated cells (35–100 × 20–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 25a) 25–40 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 10–20 × 8–15 μm. Basidia (Fig. 25a) 50–70 × 13–17 μm, clavate, 4-spored; sterigmata 4–7 μm long; basal septa lacking clamps. Basidiospores (Fig. 25b) [120/4/4] (9.5–) 10.0–12.0 (–12.5) × (8.0–) 9.0–11.0 μm, Q = 1.0–1.24 (–1.41), Qm = 1.12 ± 0.07, globose, subglobose to broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (15–55 × 15–40 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 50–65 μm thick; upper layer (15–25 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless to yellowish, filamentous hyphae 2–6 μm wide; lower layer (25–50 μm thick) composed of radially arranged, filamentous hyphae 3–7 μm wide, colorless to yellowish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 25c, d) composed of two parts intergrading to each other. Outer part of volval remnants on stipe base (Fig. 25c) composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant, 3–9 μm wide, colorless to yellowish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to fairly abundant, subglobose, ellipsoid to fusiform, 40–75 × 15–50 μm, colorless to yellowish, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Inner part of volval remnants on stipe base (Fig. 25d) composed of longitudinally arranged elements: filamentous hyphae very abundant, 3–9 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, subglobose, ellipsoid to fusiform, 35–70 × 20–60 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of outer part, but with more abundant, filamentous hyphae; inner surface gelatinized and similar to structure of inner part. Stipe trama composed of longitudinally arranged, clavate terminal cells, 45–380 × 15–50 μm; filamentous hyphae scattered to abundant, 2–12 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer to autumn.

Distribution: Known from central and southwestern China.

Additional specimens examined: CHINA. ANHUI PROVINCE: Huoshan County, in a mixed forest with Fagaceae and Pinaceae, altitude 520 m, 27 July 2013, Qi Zhao 1824 (HKAS 80934); Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 1171 m, 21 July 2017, Yan-Jia Hao 1516 (HKAS 100608). YUNNAN PROVINCE: Lijiang, in a forest of Pinus yunnanensis, altitude 2200 m, 4 August 2011, Qing Cai 543 (HKAS 70139).

Commentary: Amanita pallidozonata is well characterized by its dark brown to brown pileus with a distinctly pale colored ring-like zone at the proximal end of the marginal striations, and globose, subglobose to broadly ellipsoid basidiospores (10.0–12.0 × 9.0–11.0 μm).

Our phylogenetic analysis (Fig. 1b) indicated that A. pallidozonata is sister to A. pallidocarnea. However, A. pallidocarnea can be easily distinguished from A. pallidozonata by its red to pinkish lamellae, a pinkish stipe covered with minute, concolorous squamules and associations with broad-leaved trees (Boedijn 1951; Yang 2001, 2005, 2015).

The pale, ring-like zone at the proximal end of the marginal striations and globose to subglobose basidiospores of A. pallidozonata are reminiscent of A. umbrinolutea, but the latter has a darker colored basidioma, and scarce inflated cells in the volval remnants on the stipe base (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015; this study).

Amanita zonata, a novel species described in this study, also has a ring-like zone at the proximal end of the marginal striations and could be confused with A. pallidozonata. However, A. zonata has relatively smaller basidiospores (9.0–10.5 × 8.5–10.0 μm) and it is found in tropical forests with Fagaceae.

The Chinese A. brunneofuliginea resembles A. pallidozonata in that they share a brown-colored pileus. However, A. brunneofuliginea has a relatively larger basidioma, a brown pileus lacking a ring-like zone at the proximal end of the marginal striations, and abundant inflated cells in the outer part of the volva on the stipe base (Yang 1997, 2005, 2015).

68. Amanita pseudovaginata Hongo, Mem. Fac. Liberal Arts Shiga Univ. Nat. Sci. 33: 39 (1983).

Pileus 3–6 cm in diam., gray (2C1–2) to grayish (1B1–2); volval remnants on pileus absent, or sometimes covered with felted, patches to verrucose, white (1A1) to dirty white (2B1) volval remnants; margin striate (0.2–0.4 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe white (1A1) to dirty white (2B1); basal bulb absent; volval remnants on stipe saccate, 1.5–2 cm high × 1–2.5 cm wide, white (1A1) to dirty white (2B1). Annulus absent.

Basidia 40–65 × 15–17 μm, clavate, 4-spored. Basidiospores [180/9/9] (8.5–) 9.5–12.5 (–14.0) × (7.0–) 8.0–10.5 (–11.0) μm, Q = (1.01–) 1.05–1.33 (–1.43), Qm = 1.19 ± 0.09, subglobose to broadly ellipsoid, inamyloid. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forest with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central, eastern, northwestern, southern and southwestern China (Yang 1997, 2005, 2015; this study), India (Bhatt et al. 2003) and Japan (Hongo 1983; Yang and Doi 1999; Imazeki et al. 2011).

Specimens examined: CHINA. SICHUAN PROVINCE: Xichang, Luojishan, in a pine forest dominated by Pinus yunnanensis, altitude 2000 m, 28 July 2012, Yan-Jia Hao 654 (HKAS 76312). YUNNAN PROVINCE: Longling, xxx, altitude 1800 m, 23 August 2014, Pan-Meng Wang 224 (HKAS 95305); Ninglang County, in a forest of Pinus yunnanensis, altitude 2300 m, 6 August 2011, Qing Cai 574 (HKAS 70170); Jianchuan County, in a mixed forest with Fagaceae and Pinaceae, altitude 2300 m, 29 August 2009, Gang Wu 190 (HKAS 57722); Kunming, Heilongtan, in a subtropical forest of Pinus yunnanensis, altitude 1900 m, 23 June 2000, Zhu L. Yang 2887 (HKAS 35973); Lanping County, in a pine forest dominated by Pinus yunnanensis, altitude 2400 m, 16 August 2011, Yan-Jia Hao 552 (HKAS 71661); Lijiang, Gucheng, in a forest of Pinus yunnanensis, altitude 2200 m, 4 August 2011, Qing Cai 542 (HKAS 70138); Yulong Naxi Autonomous County, in a pine forest dominated by Pinus yunnanensis, altitude 2655 m, 31 July 2011, Li-Ping Tang 1372 (HKAS 69833). JAPAN: HONSHU: Kyoto, Uji-city, Ikeno-o, in a mixed forest with Pinus and Quercus, 4 August 1981, T. Hongo 6134 (HOLOTYPE, TNS-F-237282).

Commentary: Amanita pseudovaginata was described from Japan by Hongo (1983), and subsequently reported from China and India (Yang 1997, 2005, 2015; Bhatt et al. 2003). In our multi-locus phylogenetic analysis (Fig. 1b), A. pseudovaginata was found to be related to A. brunneofuliginea and A. aff. brunneofuliginea. However, A. brunneofuliginea can be easily distinguished from A. pseudovaginata by its dark brown pileus, a saccate volva on the stipe base with its outer surface covered with brown squamules and its subalpine distribution. Collections of A. aff. brunneofuliginea listed under A. brunneofuliginea have relatively broader basidiospores measuring [60/3/3] 11.0–13.0 × 10.0–12.0 (–12.5) μm, Q = 1.0–1.1, Qm = 1.05 ± 0.03, and are associated with trees of the genera Abies, Picea and Fagus. For detailed descriptions, comparisons with similar species, line drawings and images of A. pseudovaginata see Yang (1997, 2005, 2015).

69. Amanita retenta Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22h, i, 26.
Fig. 26

Microscopic features of Amanita retenta. a Hymenium and subhymenium (HKAS 54604); b basidiospores (TYPE, HKAS 70020); c longitudinal section of outer part of volval remnants on stipe base (TYPE, HKAS 70020); d longitudinal section of inner part of volval remnants on stipe base (TYPE, HKAS 70020). Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825029

Etymology: retenta, form retentus = retained, referring to the volval remnants on pileus often retained as large, white patches slightly attached on pileus of this species.

Type: CHINA. YUNNAN PROVINCE: Lijiang, in a mixed forest with Fagaceae and Pinaceae, altitude 2438 m, 28 July 2011, Qing Cai 424 (TYPE, HKAS 70020, GenBank Acc. No.: ITS = MH508543, nrLSU = MH486802, tef1-α  = MH509028, rpb2 = MH486215, β-tubulin  = MH485721).

Basidioma (Fig. 22h, i) small to medium-sized. Pileus 3–7 cm diam., plano-convex to applanate, lacking an umbo or depression at center, dirty white (2B1), grayish (1B1–2), gray (2C1–3), gray-brown (3C1–4) to brown (4B3–5), occasionally pure white (1A1) over entire disk; volval remnants often persistent as large, white (1A1) patches slightly attached on pileus, or sometimes absent; margin striate (0.15–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edge white (1A1); lamellulae truncate, plentiful. Stipe 7–11 cm long × 0.4–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1B1), dirty white (2B1) to brownish (1B1–3), densely covered with minute, concolorous, snakeskin-shaped squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 1–4 cm high × 1–2 cm wide, membranous, both surface white (1B1) to dirty white (2B1), with limbus internus placed on inside of volval limb or at point of attachment between stipe and volval limb, and upper part of volva sometimes easily cracking into large patches slightly attached on lower part of stipe. Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant, subglobose, fusiform, ellipsoid to clavate inflated cells (30–110 × 20–25 μm); filamentous hyphae abundant, 2–7 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid to elongate inflated cells (30–70 × 15–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 5–8 μm wide. Subhymenium (Fig. 26a) 40–55 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 8–15 × 8–15 μm. Basidia (Fig. 26a) 50–65 × 15–19 μm, clavate, 4-spored; sterigmata 5–7 μm long; basal septa lacking clamps. Basidiospores (Fig. 26b) [140/9/9] 10.0–12.0 (–13.0) × (9.0–) 9.5–11.5 (–12.0) μm, Q = 1.00–1.16 (–1.2), Qm = 1.08 ± 0.04, globose to subglobose, rarely broadly ellipsoid, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (10–60 × 10–40 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 100–150 μm thick; upper layer (25–60 μm thick) gelatinized, composed of radially arranged to interwoven, thin-walled, colorless to yellowish, filamentous hyphae 2–5 μm wide; lower layer (50–110 μm thick) composed of radially arranged, filamentous hyphae 3–6 μm wide, yellow-brown; vascular hyphae scarce. Volval remnants on stipe base (Fig. 26c, d) dominantly composed of two parts intergrading to each other. Outer part of volval remnants on stipe base (Fig. 26c) composed of more or less longitudinally arranged elements: filamentous hyphae abundant to very abundant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to nearly dominant, subglobose, ellipsoid to fusiform, 35–80 × 20–55 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Inner part of volval remnants on stipe base (Fig. 26d) gelatinized, composed of longitudinally arranged elements: filamentous hyphae abundant to very abundant to dominant, 2–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, subglobose, ellipsoid to fusiform, 40–60 × 20–50 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Stipe trama composed of longitudinally arranged, clavate terminal cells, 60–300 × 15–45 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer to autumn.

Distribution: presently known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lijiang, in a mixed forest, altitude 2438 m, 28 July 2011, Qing Cai 428 (HKAS 70024); same location, in a forest with Fagaceae and Pinus yunnanensis, altitude 2415 m, 28 July 2011, Li-Ping Tang 1326 (HKAS 69787); same location, in a forest with Fagaceae and Pinus yunnanensis, altitude 3000 m, 22 July 2008, Li-Ping Tang 441 (HKAS 54672); same city, Heilongtan, in a forest of Pinus densata, altitude 1980 m, 1 September 2013, Bang Feng 1485 (HKAS 82591); same city, in a pine forest, altitude 2880 m, 18 August 2010, Xue-Tai Zhu 203 (HKAS 68379); same location, in a mixed forest, altitude 2700 m, 3 August 2011, Qing Cai 534 (HKAS 70130); same city, Xiangshan, in a mixed forest, altitude 2100 m, 19 July 2008, Li-Ping Tang 373 (HKAS 54604); same city, Yulongxueshan, in a pine forest, altitude 2800 m, 21 July 2008, Li-Ping Tang 423 (HKAS 54654).

Commentary: Amanita retenta is characterized by its small to medium-sized basidioma with a gray to gray-brown pileus often covered with large, white volval patches, globose to subglobose basidiospores (10.0–12.0. × 9.5–11.5 μm), and is found in pine or mixed forests. Because there are abundant inflated cells in the outer part of the volva, the volva can easily be broken during the development of the basidioma. Specimens of A. retenta with entirely white basidiomata have also been collected in the field.

Our phylogenetic analysis (Fig. 1b) did not uncover any species closely related to A. retenta. Amanita retenta is morphologically similar to A. nivalis, described from Europe. However, A. nivalis has a relatively shorter basidioma with its stipe ca. 4–8 cm long, and was described as a snow-bed agaric in association with Carex, Galium and Polytrichum (Gilbert 1940, 1941a; Watling 1985; Campo and Bizio 2000; Contu 2000b; Galli 2001; Neville and Poumarat 2004).

Amanita ovalispora and A. pseudovaginata are also similar to A. retenta in macroscopic appearance. However, they can be separated from A. retenta in their broadly ellipsoid to ellipsoid basidiospores (8.5–10.0 × 6.5–7.5 μm for A. ovalispora; 10.5–13.0 × 8.0–9.0 μm for A. pseudovaginata), and the inflated cells are concentrated in the inner part of the volva (Boedijn 1951; Hongo 1983; Yang 1997, 2005, 2015; Yang and Doi 1999).

The white basidioma of A. retenta could be misidentified as the European A. vaginata var. alba Gillet. For example, HKAS 82591 was erroneously treated as A. vaginata var. alba by Yang (2015) and should be a species of A. retenta based on our ITS phylogenetic analysis (Fig. S8). However, A. vaginata var. alba differs from the white basidioma of A. retenta by the absence of volval remnants on the pileus, and the irregularly distributed, brownish spots on the outer surface of the volva on the stipe base (Gilbert 1940, 1941a; Contu 2000a, b; Galli 2001).

70. Amanita shennongjiana Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22j, k, 27.
Fig. 27

Microscopic features of Amanita shennongjiana. a Hymenium and subhymenium (TYPE, HKAS 75553); b basidiospores (TYPE, HKAS 75553); c longitudinal section of outer part of volval remnants on stipe base (HKAS 75554); d longitudinal section of inner part of volval remnants on stipe base (HKAS 75554). Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825030

Etymology: shennongjia, referring to the type locality of this species.

Type: CHINA. HUBEI PROVINCE: Yichang, Shennongjia, in a forest of Quercus, altitude 1850 m, 15 July 2012, Qing Cai 799 (TYPE, HKAS 75553, GenBank Acc. No.: ITS = MH508590, nrLSU = MH486862, tef1-α  = MH509085, rpb2 = MH486270, β-tubulin  = MH485771).

Basidioma (Fig. 22j, k) small to medium-sized. Pileus 4–9 cm diam., plano-convex to applanate, umbonate at center, grayish brown (3C2–5) to brownish (1B2–4) over disk; volval remnants on pileus mostly absent, or occasionally retained as white patches; margin striate (0.4–0.5 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edges white (1A1); lamellulae truncate, plentiful. Stipe 10–16 cm long × 0.5–1.2 cm diam., slender, subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with minute, concolorous fibrils; context white (1A1), hollow in center; basal bulb absent; volva saccate, 1.5–3 cm high × 1–1.5 cm wide, membranous, easily broken into large patches during development of basidioma, both surfaces white (1A1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 20–30 μm wide, composed of abundant, subglobose, ellipsoid, elongate to clavate inflated cells (20–60 × 15–40 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform to ellipsoid inflated cells (40–60 × 10–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–6 μm wide. Subhymenium (Fig. 27a) 30–50 μm thick, with 2–3 layers of ovoid, fusiform to ellipsoid or irregularly arranged cells, 8–25 × 8–15 μm. Basidia (Fig. 27a) 45–60 × 15–18 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 27b) [80/2/2] (9.5–) 10–12 × 9–11 (–11.5) μm, Q = 1–1.12 (–1.2), Qm = 1.06 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, fusiform to ellipsoid or sphaeropedunculate, inflated cells (10–50 × 10–40 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 60–125 μm thick; upper layer (20–60 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless to yellowish, filamentous hyphae 2–7 μm wide; lower layer (40–65 μm thick) composed of radially arranged, filamentous hyphae 3–9 μm wide, yellowish to brownish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 27c, d) composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–10 μm wide, colorless to brownish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, ellipsoid, fusiform to elongate, 20–75 × 15–45 μm, colorless to brownish, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to structure of interior part; inner surface gelatinized and similar to structure of interior part. Stipe trama composed of longitudinally arranged, clavate, terminal cells, 90–430 × 15–55 μm; filamentous hyphae scattered to abundant, 2–13 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved forests with Fagaceae; basidioma occurring in summer to autumn.

Distribution: Known from central China.

Additional specimen examined: CHINA. HUBEI PROVINCE: Yichang, Shennongjia, in a forest with Quercus, altitude 1850 m, 15 July 2012, Qing Cai 800 (HKAS 75554).

Commentary: Amanita shennongjiana is distinguished by its slender basidioma, globose to subglobose basidiospores (10.0–12.0 × 9.0–11.0 μm) and its associations with broad-leaved trees. In addition, the saccate volva on the stipe base of A. shennongjiana is easily broken into large patches during the development of basidioma.

Our ITS phylogenetic analysis (Fig. S8) indicated that collection of LE〈RUS〉:217377 (ITS = KM658299) from Russia, which was submitted to NCBI as A. vaginata, should be A. shennongjiana. Phylogenetically, A. shennongjiana is closely related to A. griseoumbonata (Fig. 1b). Indeed, A. griseoumbonata is morphologically similar to A. shennongjiana and they share a slender basidioma with an umbonate pileus, globose to subglobose basidiospores and a similar structure of the volval remnants on the stipe base. However, A. griseoumbonata differs from A. shennongjiana by its darker-colored pileus and is found in coniferous or mixed forests.

The European A. vaginata var. vaginata is similar to A. shennongjiana in appearance, but A. vaginata var. vaginata has a robust basidioma, a non-umbonate pileus with relatively shorter striations on the margin and a relatively thicker volva on the stipe base (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; this study).

Amanita ovalispora and A. pseudovaginata are somewhat similar to A. shennongjiana. However, A. ovalispora has a robuster basidioma, broadly ellipsoid to ellipsoid basidiospores (8.5–10.0 × 6.5–7.5 μm), a more robust volva on the stipe base, and occurs in more southern regions (Boedijn 1951; this study). Amanita pseudovaginata possesses a relatively paler-colored pileus and subglobose to broadly ellipsoid basidiospores (10.5–13.0 × 8.0–9.0 μm) (Hongo 1983; Yang 1997, 2005, 2015; Yang and Doi 1999).

71. Amanita tenuifulva Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22l, m, 28.
Fig. 28

Microscopic features of Amanita tenuifulva. a Hymenium and subhymenium (TYPE, HKAS 87120); b basidiospores (TYPE, HKAS 87120); c longitudinal section of outer part of volval remnants on stipe base (HKAS 58877); d longitudinal section of inner part of volval remnants on stipe base (HKAS 58877). Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825031

Etymology: tenuifulva, from tenuis = slender, and fulvus = brown or yellow-brown, referring to its slender basidioma with a brown pileus, also meaning its similar appearance with Amanita fulva, but with a relatively slender basidioma.

Type: CHINA. YUNNAN PROVINCE: Longling County, Zhenan, in a forest with Fagaceae, altitude 2500 m, 31 July 2014, Xiao-Bin Liu 474 (TYPE, HKAS 87120, GenBank Acc. No.: nrLSU = MH486929, tef1-α  = MH509146, rpb2 = MH486322, β-tubulin  = MH485829).

Basidioma (Fig. 22l, m) medium-sized. Pileus 5–8 cm diam., plano-convex to applanate, often umbonate at center, dark brown (4E4–8) at center, but brown (3D5–8) to brownish (2C4–8) towards margin; volval remnants on pileus absent; margin striate (ca. 0.5 R), non-appendiculate; trama white (1A1) to cream (2A2), unchanging. Lamellae free, crowded, white (1A1) to cream (2A2); lamellulae truncate, plentiful. Stipe 10–14 cm long × 0.9–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, dirty white (2B1) to brownish (2C4–8), covered with white (1A1) to brown (3D5–8) tomentum; context white (1A1) to cream (2A2), hollow in center; basal bulb absent; volva saccate, 1.6–3.5 cm high × 1.3–2 cm wide, membranous, both surfaces white (1A1), with brown (3D5–8) tinge. Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 (–75) μm wide, composed of abundant ellipsoid to clavate inflated cells (45–90 × 10–35 μm); filamentous hyphae abundant, 2–6 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subglobose, fusiform to ellipsoid inflated cells (25–65 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 28a) 30–50 μm thick, with 2–3 layers of subglobose, fusiform, ellipsoid to irregularly arranged cells, 8–40 × 10–25 μm. Basidia (Fig. 28a) 45–60 (–80) × 15–20 μm, clavate, 4-spored; sterigmata 3–9 (–15) μm long; basal septa lacking clamps. Basidiospores (Fig. 28b) [120/3/2] 10.0–12.0 (–12.5) × (9.0–) 10.0–11.5 (–12.5) μm, Q = 1.00–1.08 (–1.11), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, ellipsoid to fusiform or sphaeropedunculate inflated cells (10–45 × 10–35 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 40–100 μm thick; upper layer (15–50 μm thick) gelatinized, composed of interwoven to radially arranged, thin-walled, colorless to brownish, filamentous hyphae 2–5 μm wide; lower layer (25–50 μm thick) composed of radially arranged, filamentous hyphae 2–8 μm wide, colorless to brownish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 28c, d) composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–13 μm wide, colorless to yellowish brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant, becoming abundant toward inner part, subglobose, subfusiform to ellipsoid or sometimes clavate, 25–80 × 25–50 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base similar to interior part, but with more abundant filamentous hyphae; inner surface gelatinized and similar to structure of interior part. Stipe trama composed of longitudinally arranged, clavate terminal cells, 60–420 × 15–50 μm; filamentous hyphae scattered to abundant, 2–12 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimen examined: CHINA. YUNNAN PROVINCE: Yulong Naxi Autonomous County, in a broad-leaved forest with Fagaceae, altitude 3000 m, 2 September 2009, Qing Cai 212 (HKAS 58877).

Specimens of A. fulva examined: AUSTRIA. VORARLBERG: Kleinwalsertal, 27 September 2016, Zhu L. Yang 5930 (HKAS 96168). GERMANY. FREE STATE OF BAVARIA: Bayerisches Wald Nat. Pk., 18 October 1987, N. Arnold (2) (L). FRANCE: detailed collection information unknown, J. P. Xu 44 (HKAS 101420). U. K. ENGLAND: Cumbria, Ambleside, 10 September 1988, Aaron Norarevian & Geoffrey G. Kibby s.n. [Tulloss 9-10-88-B] (RET 108-9); Dorset, Beaulieu, New Forest, 13 September 1988, B. McAdoo, R. Phillips & R. E. Tulloss 9-13-88-B (RET 108-8). SCOTLAND: Glen Affric, Caledonian Pinewood Res., 5 September 1988, Aaron Norarevian s.n. [Tulloss 9-5-88-C] (RET 107-4).

Commentary: Amanita tenuifulva is characterized by its slender basidioma, a brown pileus with long striations on the margin, globose to subglobose basidiospores (10–12 × 10–11.5 μm) and a volva with fairly abundant to abundant inflated cells in its inner part.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. tenuifulva is closely related to A. fulva, a species described from Europe. Indeed, A. tenuifulva is similar to A. fulva in their similarly colored basidioma. We have examined collections of A. fulva from Europe: the basidiospores were [145/6/4] (9.5–) 10.0–14.0 (–17.0) × (9.0–) 9.5–13.5 (–15.5) μm, Q = 1.00–1.12 (–1.17), Qm = 1.06 ± 0.03, globose to subglobose, inamyloid; volval remnants on the stipe base were composed longitudinally of two parts: inflated cells were dominant in the outer part, but became scarce towards the inner part. Based on the literature and examinations of materials, A. fulva differs from A. tenuifulva in its relatively robust basidioma with a proportionally shorter stipe, and the structure of the volval remnants on the stipe base (Contu 2000a, b; Massart 2000; Yang et al. 2004; Yang 2005, 2015).

The Chinese species, A. orientifulva, also has a brown basidioma. However, A. orientifulva differs from A. tenuifulva in its relatively robuster basidioma, larger basidiospores (10.0–14.0 × 9.5–13.0 μm) and scarce inflated cells in the outer part of the volval remnants on the stipe base (Yang et al. 2004; Yang 2005, 2015).

72. Amanita tomentosivolva Zhu L. Yang, Bibl. Mycol. 170: 107 (1997).

Pileus ca. 5.5 cm in diam., umbonate, brown (2D1–3) to grayish (1B1, 1C1); volval remnants on pileus absent; margin striate (ca. 0.3 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe white (1A1), covered with snakeskin-shaped, gray (3C1) squamules; basal bulb absent; volval remnants on stipe saccate, 4.5 cm high × 1.5 cm wide, outer surface covered with tomentose to felted or verrucose, gray (3C1) squamules. Annulus absent.

Basidia 37–55 × 13–17 μm, clavate, 4-spored. Basidiospores [30/1/1] (11.0–) 11.5–14.0 × (10.5–) 11.0–13.5 μm, Q = (1.0–) 1.02–1.07 (–1.1), Qm = 1.04 ± 0.02, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Volval remnants on stipe base composed of two layers intergrading to each other. Tomentose to felted or verrucose squamules on outer part composed of abundant filamentous hyphae and very abundant inflated cells. Inner layer composed of very abundant to dominant filamentous hyphae and scarce inflated cells. Clamps absent in all parts of basidioma.

Habitat: In a subtropical broad-leaved forest with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study).

Specimen examined: CHINA. YUNNAN PROVINCE: Pingbian County, in a subtropical forest with Fagaceae, altitude 1900 m, 4 July 1992, Zhu L. Yang 1889 (HOLOTYPE, HKAS 32504).

Commentary: Amanita tomentosivolva seems to be rare and was described from China by Yang (1997). It is well-characterized by the tomentose to felted or verrucose squamules on the outer surface of the saccate volva on the stipe base. To date, only the type is available for study. Unfortunately, the phylogenetic relationships of A. tomentosivolva are still unknown due to lack of molecular data. For detailed descriptions, comparisons with similar species, line drawings and images of A. tomentosivolva see Yang (1997, 2005, 2015).

73. Amanita umbrinolutea (Gillet) Bataille, Bull. Soc. mycol. Fr. 26: 139 (1910).

Basionym: Amanita inaurata var. umbrinolutea Gillet, Hyménomycètes (Alençon): 42 (1874).

Synonym: Amanita atrofusca Zhu L. Yang, Bibl. Mycol. 170: 73 (1997), syn. nov.

Basidioma medium-sized, sometimes large. Pileus 5–10 cm diam., plano-convex to applanate, umbonate at center, dark gray (4F5–8), gray (1E1–3), gray-brown (2E1–4) to brown-yellow (3E5–8); volval remnants on pileus mostly absent; margin striate (0.2–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to dirty white (2B1); lamellar edges brownish (3D3–5) to gray-brown (2E1–4); lamellulae truncate, plentiful. Stipe 10–20 cm long × 0.8–2.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, dirty white (2B1), covered with gray (1E1–3), dark gray (4F5–8), brown (2E1–4) to dark brown (3F6–8) squamules; context white (1A1), hollow in center; basal bulb absent; volva saccate, 3–5 cm high × 1.5–3 cm wide, membranous, outer surface dirty white (2B1), grayish (3C1) to brownish (3D3–5), covered with brownish (3D3–5) to brown (2E1–4) patches, inner surface dirty white (2B1) to brownish (3D3–5). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–40 μm wide, composed of abundant, fusiform inflated cells (50–130 × 12–30 μm); filamentous hyphae abundant, 2–7 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, fusiform to ellipsoid inflated cells (40–55 × 12–25 μm), diverging at an angle of ca. 30° to 60° to mediostratum; filamentous hyphae abundant and 3–8 μm wide. Subhymenium 30–50 μm thick, with 2–4 layers of subglobose, ovoid to ellipsoid or irregularly arranged cells, 10–25 × 10–20 μm. Basidia 50–90 × 14–23 μm, clavate, 4-spored; sterigmata 5–8 μm long; basal septa lacking clamps. Basidiospores [100/10/9] (9.0–) 10.0–13.0 (–14.0) × (8.5–) 9.5–12.0 (–12.5) μm, Q  = 1.0–1.15 (–1.24), Qm = 1.08 ± 0.05, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, fusiform to ellipsoid or sphaeropedunculate inflated cells (35–80 × 25–40 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–4 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 60–120 μm thick; upper layer (20–90 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless, filamentous hyphae 1–3 μm wide; lower layer (20–80 μm thick) composed of radially arranged, filamentous hyphae 2–7 μm wide, brownish; vascular hyphae scarce. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–10 μm wide, colorless to brownish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, ellipsoid to fusiform, 35–100 × 18–50 μm, colorless to brownish, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base dominantly composed of very abundant to dominant filamentous hyphae, and scarce to not easily observed inflated cells; inner surface gelatinized and composed of very abundant filamentous hyphae. Stipe trama composed of longitudinally arranged, clavate terminal cells, 150–330 × 25–50 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered common in alpine to subalpine forests with Pinaceae; basidioma occurring in summer to autumn.

Distribution: Known from northern, northwestern, southern and southwestern China (Yang 2005, 2015; this study), Europe (Consiglio 2000; Rava 2000; Tulloss et al. 2001; Neville and Poumarat 2009), India (Bhatt et al. 2003), Pakistan (Tulloss et al. 2001) and the Islamic Republic of Iran (Saber 1997).

Specimens examined: SICHUAN PROVINCE: Batang County, in a subalpine forest, altitude 3700 m, 7 August 2014, Kuan Zhao 706 (HKAS 89637); same location, in a subalpine forest of Picea, altitude 3700 m, 7 August 2014, Bang Feng 1703 (HKAS 94089); Hongyuan County, in a subalpine forest of Abies, altitude 3600 m, 14 July 1991, Ming-Sheng Yuan 1377 (HKAS 24930); Kangding County, in a subalpine forest, altitude 3500 m, 8 September 2016, KD-LJW 113 (HKAS 97906); Luhuo County, in a subalpine forest, altitude 3600 m, 15 July 2014, Kuan Zhao 538 (HKAS 89201). TIBET AUTONOMOUS REGION: Changdu, Laduo, in a subalpine forest of Picea, altitude 3800 m, 29 July 2009, Zhu L. Yang 5390 (HKAS 57932). YUNNAN PROVINCE: Shangri-La, Daxueshan, in a subalpine forest dominated by Abies and Picea, altitude 4000 m, 23 August 2000, Zhu L. Yang 2986 (HKAS 36610). GERMANY: FREE STATE OF BAVARIA: Balderschwang, altitude 1160 m, 6 September 2011, KR 10656 (MB-000658).

Commentary: In our ITS phylogenetic analysis (Fig. S8), the samples of A. atrofusca from China clustered with those of the European A. umbrinolutea. Although there are some subtle morphological differences between the Chinese populations and the European ones (Yang 1997, 2005, 2015), these may be caused by different environmental conditions. Therefore, A. atrofusca is treated here as a synonym of A. umbrinolutea.

Amanita umbrinolutea is phylogenetically related to A. pachycolea D. E. Stuntz and Amanita sp. (Figs. 1b, S8). However, Amanita pachycolea differs from A. umbrinolutea in that its white lamellae are unchanging or becoming tawny to orange-brown or orange-yellow with age, and the presence of clamps (Thiers and Ammirati 1982; Thiers 1982; Jenkins 1986; Tulloss 1994). Amanita sp. [HUBEI PROVINCE: Yichang, Shennongjia, in a subalpine forest with Abies, altitude 1500 m, 10 July 2012, Qing Cai 720 (HKAS 75475)] has a yellow-brown pileus with a yellowish to brownish margin and a brownish stipe.

74. Amanita vaginata (Bull.) Lam. var. vaginata, Encycl. Méth. Bot. (Paris) 1(1): 109 (1783).

Basionym: Agaricus vaginatus Bull., Herb. Fr. (Paris) 3: tab. 98 (1783) [1782-83].

Pileus 3–8 cm in diam., somewhat umbonate, gray (1D1–2, 1C1); volval remnants on pileus absent or retained as white (1A1) patches; margin striate (0.1–0.3 R), non-appendiculate. Lamellae white (1A1); lamellulae truncate. Stipe 5–10 cm long × 0.5–1.5 cm diam., white (1A1) to dirty white (2B1), glabrous or covered with fibrous, grayish (3B1) to brownish (1C2–3) fibrils; basal bulb absent; volval remnants on stipe base saccate, outer surface white (1A1) to dirty white (2B1), inner surface white (1A1). Annulus absent.

Basidia 48–60 × 13–18 μm, clavate, 4-spored. Basidiospores [205/20/20] (9.0–) 9.5–11.0 (–14.0) × (8.5–) 9.0–10.5 (–13.5) μm, Q = 1.0–1.08 (–1.12), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from Asia (Imai 1933, 1938; Tulloss et al. 2001; Yang 2005; Imazeki et al. 2011; Kim et al. 2013b), North America (Coker 1917; Thiers 1982; Jenkins 1986; Tulloss et al. 1995) and Europe (Horak 1968; Consiglio 2000).

Specimens examined: CHINA. BEIJING: Donglingshan, altitude unknown, 19 August 1998, Hua-An Wen & Shu-Xiao Sun 98294 (HMAS 75237). GUANGDONG PROVINCE: Fengkai County, Heishiding, in a forest dominated by Fagaceae, altitude unknown, 15 June 1986, Yuan-Fang Liu et al. (GDGM 10857); Qujiang, altitude unknown, 10 September 1984, Tai-Hui Li (GDGM 8146). HAINAN PROVINCE: Changjiang County, Bawangling, in a tropical to subtropical forest, altitude unknown, 18 May 1988, Huan-Qiang Chen (GDGM 14480); Ledong County, Jianfengling, in a tropical to subtropical forest, altitude unknown, 15 May 1988, Guo-Yang Zhen (GDGM 14410). HUNAN PROVINCE: Changsha, altitude unknown, collector unknown (MHHNU 4189). JIANGSU PROVINCE: Nanjing, altitude unknown, 14 June 1937, S. C. Li 168 (BPI 751472, CUP-CH 1157). TAIWAN PROVINCE: Nantou County, Huisunlinchang, altitude ca. 700 m, 25 October 2001, Jian-Ming Chen 3100 (HKAS 38794); Nantou County, Riyuetan, altitude unknown, 4 September 2002, Jian-Ming Chen 3568 (HKAS 41319). SICHUAN PROVINCE: Daofu County, altitude unknown, 9 August 1988, Xian-Cai Dai & Tai-Hui Li (GDGM GZ0154); Xichang, altitude ca. 2000 m, 13 June 1971, Shi-Xuan Wu et al. 19c (HMAS 35974c); Yajiang County, altitude unknown, 6–8 August 1983, Jing-Jun Sun & Hua-An Wen 464 (HMAS 51048); same location, altitude unknown, 6–8 August 1983, Jing-Jun Sun & Hua-An Wen 608 (HMAS 50826); same location, altitude unknown, 6–8 August 1983, Jing-Jun Sun & Hua-An Wen 621 (HMAS 50992). YUNNAN PROVINCE; Binchuan County, Jizushan, altitude unknown, 7 August 1989, Yu-Chen Zong & Yu Li 151 (HMAS 54117); Lijiang, Yulongxueshan, altitude 3200 m, 2 August 1995, Zhu L. Yang 2128 (HKAS 29503); Shangri-La, altitude unknown, 24 July 1986, Yu Li L50 (HMAS 59825); same location, altitude unknown, 24 July 1986, Yu Li 66 (HMAS 59781). NETHERLANDS. UTRECHT PROVINCE: Baarn, Eembrugge, 10 October 1993, H. A. v. d. Aa s.n. (L).

Commentary: The concept of A. vaginata var. vaginata remains rudimentary because no nomenclatural type has been assigned in order to delimit it precisely. Weiß et al. (1998) provided an nrLSU sequence of A. vaginata var. vaginata based on a specimen, H. A. v. d. Aa s.n. (L), and we follow Weiß’s treatment here (Fig. 2). We have examined the collection H. A. v. d. Aa s.n. (L), and its basidiospores were [70/2/1] (10–) 10.5–13.0 (–14.0) × (9.0–) 9.5–11.5 (–12.5) μm, Q = 1.02–1.16 (–1.25), Qm = 1.11 ± 0.05. It is generally recognized that A. vaginata var. vaginata is characterized by a gray pileus with striations at the margin, a white stipe lacking an annulus and globose to subglobose basidiospores [9.0–13.0 (–14.0) μm] (Lange 1935; Huijsman 1959; Bas 1967; Horak 1968; Jenkins 1986; Breitenbach and Kränzlin 1995; Tulloss et al. 1995). Based on these characters, we treated the Chinese collections listed in Yang (2005) as A. vaginata var. vaginata for the time being. Unfortunately, we were unable to generate DNA sequences from these Chinese collections.

Our multi-locus phylogenetic analysis (Fig. 1b) did not suggest any species closely related to A. vaginata var. vaginata. In addition, HKAS 56255, treated as A. vaginata var. vaginata by Yang (2015), should be A. battarrae (Fig. 1b). For detailed descriptions, comparisons with similar species, line drawings and images of the Chinese A. vaginata var. vaginata see Yang (2005).

75. Amanita vaginata var. alba (De Seynes) Gillet, Hyménomycètes (Alençon): 51 (1874) [1878].

Basionym: Agaricus vaginatus var. albus De Seynes, Essai Fl. mycol. Montpellier: 105 (1863).

Amanita vaginata var. alba only differs from A. vaginata var. vaginata by its white (1A1) basidioma, sometimes with cream (1A2) tinge.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from Asia (Yang 2005; Imazeki et al. 2011) and Europe (Gilbert 1940, 1941a; Tulloss and Moses 1995; Massart 2000).

Specimens examined: CHINA. TAIWAN PROVINCE: Taizhong, altitude unknown, 31 October 2001, Jian-Ming Chen 3187 (HKAS 38790). YUNNAN PROVINCE: Lijiang, altitude 2900 m, 30 July 2001, Zhu L. Yang 3110 (HKAS 38287).

Commentary: The Chinese collections with a white pileus similar to A. vaginata var. vaginata are treated as A. vaginata var. alba for the time being. The precise classification of A. vaginata var. alba can only be done with an assignment of a neotype of A. vaginata var. vaginata. Our phylogenetic analysis suggested that HKAS 82591, treated as A. vaginata var. alba by Yang (2015), should in fact be A. retenta (Fig. S8). For detailed descriptions, comparisons with similar species, line drawings of the Chinese A. vaginata var. alba see Yang (2005).

76. Amanita verrucosivolva Zhu L. Yang, Bibl. Mycol. 170: 111 (1997).

Pileus 7–14 cm in diam., umbonate, orange (5D5–8) to yellow (5B4–6) at center, but yellowish (4A3–6) at margin; volval remnants on pileus absent; margin striate (0.3–0.5 R), non-appendiculate. Lamellae white (1A1) to cream (1A2); lamellar edges yellowish (4A3–6); lamellulae truncate. Stipe 10–22 cm long × 1–2 cm diam., white (1A1) to cream (1A2), covered with snakeskin-shaped, yellow (5B4–6) to orange (5D5–8) squamules; basal bulb absent; volval remnants on stipe base saccate, 4–8 cm high × 2–4 cm wide, with its outer surface covered with verrucose, brown (4D5–7) to brownish (4C3–5) squamules. Annulus absent.

Basidia 50–75 × 13–21 μm, clavate, 4-spored. Basidiospores [220/7/6] (9.0–) 10.0–12.5 (–14.0) × (8.5–) 9.0–12.0 (–14.0) μm, Q = 1.0–1.08 (–1.28), Qm = 1.04 ± 0.03, globose to subglobose, inamyloid. Volval remnants on stipe base composed of two parts intergrading to each other. Verrucose squamules on outer part composed of very abundant inflated cells and abundant filamentous hyphae. Inner part composed of very abundant filamentous hyphae and scarce inflated cells. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical broad-leaved forests (e.g. Castanopsis, Lithocarpus and Magnolia); basidioma occurring in summer and autumn.

Distribution: Known from central, southern and southwestern China (Yang 1997, 2005, 2015; this study).

Specimens examined: CHINA. GUANGXI PROVINCE: Guilin, Maoer Mountain National Nature Reserve, in a broad-leaved forest with Fagaceae, altitude 1900 m, 24 July 2012, Qing Cai 874 (HKAS 75608). HUBEI PROVINCE: Yichang, Shennongjia, altitude unknown, 8 August 1984, Xiao-Qing Zhang and Shu-Xiao Sun 217 (HMAS 53760). YUNNAN PROVINCE: Baoshan, Gaoligongshan, in a broad-leaved forest with Fagaceae, altitude 1600 m, 10 August 2014, Xiao-Bin Liu 601 (HKAS 87156); Longling County, Zhenan, Luoboyakou, in a broad-leaved forest with Fagaceae, altitude 1900 m, 30 July 2014, Xiao-Bin Liu 460 (HKAS 87106); same location, in a broad-leaved forest with Fagaceae, altitude 1900 m, 30 July 2014, Yang-Yang Cui 166 (HKAS 83810); Maguan County, in a subtropical broad-leaved forest with Fagaceae, altitude 2000 m, 29 June 1992, Zhu L. Yang 1815 (HOLOTYPE, HKAS 28253).

Commentary: Amanita verrucosivolva was described from China by Yang (1997). Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. verrucosivolva is related to A. aff. fulva (HKAS 29518). However, the collection of A. aff. fulva has a pileus with a dark brown center and a gray-brown to brownish margin, white lamellar edges, a dirty white, brownish to grayish stipe without distinct squamules, a white, saccate volva with its outer surface covered with brownish pots. For detailed descriptions, comparisons with similar species, line drawings and images of A. verrucosivolva see Yang (1997, 2005, 2015).

77. Amanita zonata Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 22n, o, 29.
Fig. 29

Microscopic features of Amanita zonata. a Hymenium and subhymenium (TYPE, HKAS 97240); b basidiospores (TYPE, HKAS 97240); c longitudinal section of outer surface of volval remnants on stipe base (HKAS 97707); d longitudinal section of interior part of volval remnants on stipe base (HKAS 97707); e Longitudinal section of inner surface of volval remnants on stipe base (HKAS 97707). Bars: ab = 10 μm, c–e = 40 μm

MycoBank: MB 825034

Etymology: zonata, from zonatus = zoned, referring to its pileus with a dark-colored ring-like zone at proximal end of marginal striations.

Type: CHINA. YUNNAN PROVINCE: Lancang County, in a forest dominated by Fagaceae, altitude 1200 m, 26 September 2016, LC-Ding 29 (TYPE, HKAS 97240, GenBank Acc. No.: nrLSU = MH486959, tef1-α  = MH509179, rpb2 = MH486352).

Basidioma (Fig. 22n, o) medium-sized. Pileus 5–8 cm diam., applanate, gray-brown (1D5–8), brown (2C4–7) to dark gray (1E3–6) at center, but becoming gray (2B1, 3C1–2) to gray-brown (1D5–8) towards margin, often with an indistinctly dark-colored ring-like zone at proximal end of marginal striations; volval remnants on pileus absent; margin striate (0.15–0.3 R), non-appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1); lamellar edges white (1A1); lamellulae truncate, plentiful. Stipe 9–13 cm long × 0.4–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, dirty white (2B1), brownish (1B1–3) to gray-brown (1D5–8), densely covered with grayish (3B1) to brownish (1B1–3) squamules; context white (1A1) to brownish (1B1–3), hollow in center; basal bulb absent; volva saccate, 2–3.5 cm high × 1.5–2 cm wide, membranous, outer surface white (1A1), sometimes with brown (2C4–7) tinge, inner surface white (1B1) to dirty white (2B1). Annulus absent. Odor indistinct.

Lamellar trama bilateral. Mediostratum 30–60 μm wide, composed of abundant subglobose, fusiform to ellipsoid inflated cells (35–110 × 35–50 μm); filamentous hyphae abundant, 3–5 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant, ellipsoid, fusiform to elongate inflated cells (45–70 × 15–40 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 5–10 μm wide. Subhymenium (Fig. 29a) 35–45 μm thick, with 2–3 layers of subglobose to ellipsoid or irregularly arranged cells, 13–30 × 12–20 μm. Basidia (Fig. 29a) 45–60 × 12–15 μm, clavate, 4-spored; sterigmata 4–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 29b) [120/5/5] 9.0–10.5 (–12.0) × (8.0–) 8.5–10.0 (–11.0) μm, Q = 1.00–1.11 (–1.14), Qm = 1.05 ± 0.04, globose to subglobose, inamyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose, fusiform to ellipsoid or sphaeropedunculate inflated cells (10–35 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 75–125 μm thick; upper layer (20–60 μm thick) gelatinized, composed of radially arranged, thin-walled, colorless, filamentous hyphae 2–5 μm wide; lower layer (40–60 μm thick) composed of radially arranged, filamentous hyphae 3–7 μm wide, colorless to gray brownish; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 29d) composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 2–11 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce, subglobose, ellipsoid to fusiform, 35–60 × 15–40 μm, colorless, thin-walled, mostly terminal or sometimes in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base (Fig. 29c) similar to structure of interior part, but with more abundant, filamentous hyphae; inner surface of volval remnants on stipe base (Fig. 29e) gelatinized, composed of very abundant, filamentous hyphae, and scattered to fairly abundant, ellipsoid, inflated cells. Stipe trama composed of longitudinally arranged, clavate, terminal cells, 100–380 × 12–40 μm; filamentous hyphae scattered to abundant, 3–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical forests dominated by Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lancang County, in a forest dominated by Fagaceae, altitude 1150 m, 26 September 2016, LC-Ding 33 (HKAS 97244); same location, in a forest dominated by Fagaceae, altitude 1370 m, 26 September 2016, LC-LJW 201 (HKAS 97705); same location, in a forest dominated by Fagaceae, altitude 1150 m, 26 September 2016, LC-LJW 202 (HKAS 97706); same location, in a forest dominated by Fagaceae, altitude 1150 m, 26 September 2016, LC-LJW 203 (HKAS 97707).

Commentary: Amanita zonata is characterized by its gray to gray-brown pileus often with an indistinctly dark-colored ring-like zone at the proximal end of the marginal striations, globose to subglobose basidiospores (9.0–10.5 × 8.5–10.0 μm), a saccate volva with dominant filamentous hyphae, and associations with trees of the family Fagaceae in tropical forests.

The species closely related to A. zonata were not suggested by our multi-locus phylogenetic analysis (Fig. 1b). Amanita zonata can be confused with A. umbrinolutea because of the gray-brown pileus with an indistinct dark-colored ring-like zone at the proximal end of the marginal striations, and similar basidiospores. However, A. umbrinolutea, a species described from Europe and also found in China, has a darker basidioma, brownish to gray-brown lamellar edges, obvious brown spots on the outer surface of the volva, and relatively larger basidiospores (10.0–13.0 × 9.5–12.0 μm) (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015; this study). Furthermore, A. umbrinolutea is dominant in temperate regions (Gilbert 1940, 1941a; Contu 2000a, b; Consiglio 2000; Massart 2000; Galli 2001; Neville and Poumarat 2004; Yang 2005, 2015; this study).

Amanita ovalispora, a species described from Indonesia, is similar to A. zonata in appearance and habitats, but the former can be distinguished from the latter by the grayer to darker gray pileus lacking an obvious dark-colored ring-like zone at the proximal end of the marginal striations, and broadly ellipsoid to ellipsoid basidiospores (8.5–10.0 × 6.5–7.5 μm) (Boedijn 1951; Yang 1997, 2005, 2015).

In China, A. brunneofuliginea is somewhat similar to A. zonata in the gray-brown pileus. However, A. brunneofuliginea can be easily distinguished from A. zonata by its darker pileus lacking a ring-like zone at the proximal end of the marginal striations, brown squamules on the outer surface of the volva, subglobose to broadly ellipsoid basidiospores (10.5–13.0 × 9.5–12.0 μm), and habitat in plantations in subalpine forests (Yang 1997, 2005, 2015).

Amanita subgen. Amanitina (E. J. Gilbert) E. J. Gilbert, Not. Amanites XXX: 3 (1941).

Type: Amanita phalloides (Vaill.: Fr.) Link, Handbuch zur Erkennung der Nutzbarsten und am Häufigsten Vorkommenden Gewächse 3: 272 (1833).

Diagnosis: Basidioma mostly agaricoid, occasionally sequestrate. When basidioma agaricoid, pileus margin non-striate; lamellulae attenuate; basidiospores amyloid; clamps present or absent in all parts of basidioma. When basidioma sequestrate, basidia thick-walled; basidiospores amyloid, very rarely inamyloid; clamps absent in all parts of basidioma.

Description: Basidioma mostly agaricoid, rarely sequestrate. When basidioma agaricoid, pileus convex to applanate, white, dirty white, grayish, gray, brownish, brown to yellow; volval remnants on pileus present or absent; margin mostly non-striate; trama mostly white to cream, or sometimes yellow, gray, brown or pinkish, unchanging or sometimes with color change when injured. Lamellae free, white, cream, yellow, brownish, brown or greenish; lamellulae often attenuate. Stipe subcylindric or slightly tapering upwards, with apex slightly expanded; stipe base inflated or not; volval remnants on stipe base present. Annulus mostly persistent, sometimes fragile and fugacious. Lamellar trama bilateral. Subhymenium composed of 2–3 layers of subglobose to ellipsoid or irregular cells. Basidia clavate, 4-spored, rarely 2-spored. Basidiospores thin-walled, smooth, amyloid. Lamellar edge appearing as a sterile strip, composed of inflated cells and filamentous hyphae, irregularly arranged or ± running parallel to lamellar edge. Pileipellis often slightly gelatinized, composed of subradially to somewhat interwoven, thin-walled, filamentous hyphae. Volval remnants composed of abundant to very abundant, inflated cells, and scarce to fairly abundant filamentous hyphae. Stipe trama composed of longitudinally arranged, long clavate terminal cells mixed with abundant filamentous hyphae. Annulus composed of inflated cells and filamentous hyphae. Clamps present or absent in all parts of basidioma. When basidiomata sequestrate, stipitate, basidia thick-walled, basidiospores amyloid very rarely inamyloid, clamps absent.

Commentary: Vizzini et al. (2012) and Redhead et al. (2016) proposed Aspidella and Saproamanita respectively to accommodate Amanita vittadinii and its allies (A. subsect. Vittadiniae Bas), and resurrected the subgeneric name Amanitina, typified by A. phalloides, for the remaining species of A. subgen. Lepidella sensu Bas (1969), with the exception of A. subsect. Vittadinii. Our multi-locus phylogenetic analysis indicates that A. sect. Lepidella sensu Bas (1969) is polyphyletic (Fig. 1a, b). To keep the infrageneric taxonomic stability of Amanita (Tulloss et al. 2016), we follow the concept of A. subgen. Amanitina as circumscribed by Redhead et al. (2016), but retain the species complex of Amanita vittadinii as its own section, A. sect. Lepidella Corner & Bas.

Key to the sections of Amanita subgen. Amanitina

  1. 1.

    Basidioma agaricoid; basidia thin-walled; clamps present or absent; basidioma distributed worldwide and associated with Fagaceae, Pinaceae, etc……….2

     
  2. 1.

    Basidioma agaricoid or sequestrate; basidia thin- or thick-walled; clamps absent; basidioma distributed in south-western and western Australia and associated with Eucalyptus, Allocasuarina, etc………sect. Arenariae

     
  3. 2.

    Pileal margin mostly non-appendiculate; annulus membranous and not fragile, dominantly composed of filamentous hyphae……….3

     
  4. 2.

    Pileal margin mostly appendiculate; annulus membranous to fragile, dominantly composed of inflated cells……….4

     
  5. 3.

    Volval remnants often as pyramids, verrucae, cones, floccules or patches on pileal surface and base of stipe; stipe with a bulbous base………sect. Validae

     
  6. 3.

    Volval remnants on pileal surface mostly absent or leaving only flat patches, volval remnants on stipe base membranous and limbate; stipe with a bulbous base or not………sect. Phalloideae

     
  7. 4.

    Pileal margin non-striate; volval remnants fragile and often as pyramids, verrucae, cones, floccules, patches to pulverulence on pileal surface and base of stipe, sometimes pileal surface glabrous, and base of stipe with a membranous and limbate volva; lamellae lacking brown color changed wh4.en dried……….5

     
  8. 4.

    Pileal margin striate, sometimes non-striate; volval remnants on pileus often consisting of a membranous outer layer and a friable inner layer, inner layer often torn into squamules, floccules to pulverulence, volval remnants on stipe base membranous and saccate; lamellae often becoming brown when dried………sect. Amidella

     
  9. 5.

    Basidioma large to very large; base of stipe ventricose to napiform, often radicating; clamps absent………sect. Strobiliformes

     
  10. 5.

    Basidioma small, medium-sized, large to very large; base of stipe globose to subglobose, often not radicating; clamps present or not………sect. Roanokenses

     

Amanita sect. Amidella (E. J. Gilbert) Konrad & Maubl., Agaricales: 61 (1948).

Basionym: Amidella E. J. Gilbert, in Bres., Iconogr. Mycol. 27 Suppl. 1(1): 71 (1940).

Type: Amanita volvata (Peck) Lloyd, Mycol. Writ. 1(7): 9 (1898).

Diagnosis: Basidioma agaricoid; pileus white, dirty white to brownish, covered with floccose to squamulose volval remnants; lamellae white to cream-colored, becoming grayish, gray-brown, brownish to brown when dried, lamellulae truncate to nearly truncate; annulus fragile and fugacious, volva saccate at base of stipe; basidiospores amyloid and clamps absent.

Commentary: Previously, four species of this section, A. avellaneosquamosa (S. Imai) S. Imai, A. clarisquamosa (S. Imai) S. Imai, A. neoovoidea Hongo and A. rufobrunnescens W. Q. Deng & T. H. Li have been reported from China (Yang 2015; Deng et al. 2016). However, our phylogenetic data (Fig. 1b) indicate that A. neoovoidea should be transferred to A. sect. Roanokenses. We describe here four novel species, which, together with the three previously known species give a total of seven species of A. sect. Amidella recognized in China. They can be separated from each other by the size of the basidioma, the length of striations on the pileal margin, the form and the size of the basidiospores and their habitats.

Key to the species of Amanita sect. Amidella from China

  1. 1.

    Pileal margin non- or slightly striate (ca. 0.1–0.25 R) when mature……….2

     
  2. 1.

    Pileal margin with obviously striations, ca. 0.3–0.5 R………A. avellaneosquamosa

     
  3. 2.

    Pileal margin slightly striate (ca. 0.1–0.25 R) when mature; basidiospores mostly elongate with width often less than 7 μm……….3

     
  4. 2.

    Pileal margin non- or slightly striate when mature; basidiospores mostly ellipsoid with width often more than 7 μm……….6

     
  5. 3.

    Basidioma relatively larger with pileus ca. 4–10 cm in diam……….4

     
  6. 3.

    Basidioma relatively smaller with pileus ca. 3–6.5 cm in diam……….5

     
  7. 4.

    Basidioma become reddish, light brown to reddish brown when bruised or injured; dominant in broad-leaved forests with Fagaceae………A. rufobrunnescens

     
  8. 4.

    Basidioma lacking distinctly color change when bruised or injured; dominant in forests with Quercus and Abies………A. clarisquamosa

     
  9. 5.

    Basidiospores relatively larger, 10.0–12.0 × 5.5–7.0 μm, Q = 1.6–2.0, Qm = 1.81 ± 0.14; dominant in pine forests with Pinaceae………A. pinophila

     
  10. 5.

    Basidiospores relatively smaller, 9.0–11.0 × 5.0–6.0 μm, Q = 1.67–2.16, Qm = 1.92 ± 0.13; dominant in broad-leaved or mixed forests with Pinaceae and Fagaceae………A. parvicurta

     
  11. 6.

    Pileal margin non- or slightly appendiculate; basidiospores proportionally narrower, 10.0–13.0 (–15.5) × (6.0–) 6.5–8.0 (–9.0) μm, Qm = 1.65 ± 0.19; basidioma dominant in pine forests………A. brunneomaculata

     
  12. 6.

    Pileal margin strongly appendiculate; basidiospores proportionally broader, (9.0–) 10.0–12.0 (–14.0) × (6.0–) 7.0–8.5 (–9.0) μm, Qm = 1.49 ± 0.13; basidioma distributed in pine or mixed forests………A. lanigera

     

78. Amanita avellaneosquamosa (S. Imai) S. Imai, Mycol. Fl. Japan, Basidiomycetes 2: 250 (1959)

Basionym: Amanitopsis avellaneosquamosa S. Imai, Bot. Mag. (Tokyo) 47: 430 (1933).

Pileus 4–8 cm in diam., dirty white (2B1); volval remnants on pileus patchy or fibrous, brownish (4B2–4) to brown (4C2–4); margin striate (0.3–0.5 R), sometimes appendiculate. Lamellae white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate. Stipe 7–12 cm long × 0.8–2 cm diam., white (1A1) to dirty white (2B1), covered with floccose, white (1A1) to brownish (4B2–4) squamules; basal bulb absent; volval remnants on stipe base saccate, 2–6 cm long × 1.5–3 (–5) cm diam., fleshy, outer surface dirty white (2B1), inner surface white (1A1). Annulus fragile and retained as minute squamules on upper part of stipe.

Basidia 35–56 × 10–12 μm, clavate, 4-spored. Basidiospores [120/8/8] (8.0–) 9.0–11.0 (–12.5) × (4.0–) 5.0–7.0 (–7.5) μm, Q = (1.28–) 1.38–2.2 (–2.36), Qm = 1.79 ± 0.19, mostly elongate, sometimes ellipsoid or cylindrical, amyloid. Volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae abundant; inflated cells fairly abundant to abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in forests with Pinaceae or Fagaceae (e.g. Quercus, Pinus and Abies); basidioma occurring in summer and autumn.

Distribution: Known from central, eastern and southwestern China (Yang 2005, 2015; this study), India (Bhatt et al. 2003; Semwal et al. 2006a), Japan (Imai 1933, 1938; Ito 1959; Yang and Doi 1999), Republic of Korea and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 1121 m, 21 July 2017, Yan-Jia Hao 1498 (HKAS 100602). JIANGXI PROVINCE: Jiujiang, Lushan Botanical Garden, in a forest with Abies firma, altitude 1100 m, 1 September 2011, Xiao-Fei Shi 875 (HKAS 77340). TAIWAN PROVINCE: Nantou County, in a mixed forest with Quercus and Pinus, altitude 800 m, 16 September 2012, Bang Feng 1283 (HKAS 82425). YUNNAN PROVINCE: Lijiang, Yulongxueshan, in a forest of Pinus yunnanensis, altitude 2900 m, 31 July 1995, Zhu L. Yang 2106 (HKAS 29500); same location, in a forest dominated by Pinus, altitude 2580 m, 18 July 2008, Li-Ping Tang 356 (HKAS 54587). REPUBLIC OF KOREA. INCHEON: Songdo Central Park, in a mixed forest with Quercus, altitude 100 m, 16 August 2011, Xiao-Fei Shi 807 (HKAS 77339). JAPAN. HONSHU: Tokyo, Oume City, Kurosawa 3-Chôme, along right-side ridge of Kurosawa-gawa river, 25 September 1997, Y. Doi s.n. (TNS-F-237935). HOKKAIDO: Ishikari, Nopporo, altitude unknown, 17 August 1924, S. Imai (TYPE, SAPA).

Commentary: In the original description of A. avellaneosquamosa, Imai (1933) did not designate a type. Gilbert (1940) studied two collections of this taxon and designated the collection made by Imai on August 17, 1924 as the type. We have examined the type, its basidiospores were [25/1/1] 9.0–11.0 (–12.0) × (5.0–) 5.5–7.0 (–7.5) μm, Q = (1.29–) 1.44–1.88 (–2), Qm = 1.65 ± 0.16. Phylogenetically, A. avellaneosquamosa was closely related to A. parvicurta Yang-Yang Cui et al. (Fig. 1b). For comparisons between them see the commentary on A. parvicurta.

79. Amanita brunneomaculata Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 30a–c, 31.
Fig. 30

Fresh basidiomata of novel species in Amanita sect. Amidella and A. sect. Phalloideae. a–c A. brunneomaculata (a TYPE, HKAS 70032, b HKAS 71664, c-HKAS 74642); d–f A. lanigera (de TYPE, HKAS 89030, f HKAS 75018); g–h A. parvicurta (TYPE, HKAS 101215); i–l A. pinophila (i-HKAS 70165, j HKAS 68307, kl TYPE, HKAS 70167); m–o A. franzii (m TYPE, HKAS 79559, no HKAS 82832). Bars: 2 cm

Fig. 31

Microscopic features of Amanita brunneomaculata (TYPE, HKAS 70032). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base, right side shows outer surface of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825035

Etymology: brunneomaculata, from brunneus = brown, and maculatus = spotted, referring to its white pileus becoming brown spotted when mature.

Type: CHINA. YUNNAN PROVINCE: Lijiang, Jinshanxiang, in a forest of Pinus yunnanensis, altitude 2283 m, 29 July 2011, Qing Cai 436 (TYPE, HKAS 70032, GenBank Acc. No.: ITS = MH508279, nrLSU = MH486411, tef1-α  = MH508699, rpb2 = MH485893).

Basidioma (Fig. 30a–c) medium-sized. Pileus 5–9 cm diam., plano-convex to applanate, lacking an umbo or depression at center, whitish (1A1), often with brownish (1B3–5) to brown (3C2–5) spotted when mature; volval remnants on pileus floccose to patchy, whitish (1A1) to brownish (1B3–5), easily broken when mature; margin non- or slightly striate when old, appendiculate; trama white to whitish (1A1), unchanging. Lamellae free, crowded, white to off-white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate, plentiful. Stipe 5–13 cm long × 1–2.2 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1), brownish (1B3–5) to brown (3C2–5), covered with floccose, concolorous squamules; context white to whitish (1A1), soft to hollow in center; basal bulb absent; volva saccate, 1.5–5 cm long × 1.5–4 cm diam., membranous, outer surface white (1A1), brownish (4B3–5) to brown (3C2–5), inner surface brownish (4B3–5) to brown (3C2–5), 1–3 mm thick. Annulus present, pendant from attachment 3–4 cm below apex of stipe, floccose, whitish (1A1), fugacious. Odor indistinct.

Lamellar trama bilateral. Mediostratum 35–50 μm wide, composed of abundant subfusiform, ellipsoid to elongate inflated cells (40–110 × 10–40 μm); filamentous hyphae abundant, 3–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subfusiform to ellipsoid inflated cells (35–55 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 31a) 30–50 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 10–50 × 9–15 μm. Basidia (Fig. 31a) 35–50 × 9–12 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 31b) [70/4/4] 10.0–13.0 (–15.5) × (6.0–) 6.5–8.0 (–9.0) μm, Q = 1.4–1.9 (–2.25), Qm = 1.65 ± 0.19, ellipsoid to elongate, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (12–55 × 9–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 55–120 μm thick; upper layer (25–60 μm thick) gelatinized, composed of subradially to somewhat interwoven, thin-walled, colorless, filamentous hyphae 2–6 μm wide; lower layer (25–60 μm thick) composed of radially and compactly arranged, filamentous hyphae 2–10 μm wide, yellowish; vascular hyphae scarce. Volval remnants on pileus composed of irregularly arranged elements: filamentous hyphae very abundant, 2–9 μm wide, yellow to yellow-brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, ellipsoid, fusiform to clavate, 15–110 × 10–25 μm, yellow to yellow-brown, thin-walled, terminal or in chains of 2–3; vascular hyphae abundant. Interior of volval remnants on stipe base (Fig. 31c) composed of longitudinally arranged elements: filamentous hyphae very abundant, 2–8 μm wide, colorless or yellow to yellowish brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells abundant to very abundant, subglobose, fusiform to ellipsoid or pyriform, 20–70 × 10–30 μm, colorless or yellow to yellowish brown, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base (Fig. 31c) dominantly composed of very abundant filamentous hyphae, mixed with fairly abundant, pyriform, inflated cells; inner surface slightly gelatinized, similar to structure of interior part. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 70–245 × 20–45 μm; filamentous hyphae scattered to abundant, 3–8 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered under pine trees (e.g. Pinus yunnanensis); basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lanping County, in a forest of Pinus yunnanensis, altitude 2400 m, 16 August 2011, Bang Feng 1161 (HKAS 74642); same location, in a forest of Pinus yunnanensis, altitude 2400 m, 16 August 2011, Yan-Jia Hao 555 (HKAS 71664); Lijiang, in a forest of Pinus yunnanensis, altitude 2600 m, 18 August 2010, Xue-Tai Zhu 217 (HKAS 68393).

Specimens of A. volvata examined: USA. LOUISIANA, St. Tammany Parish. Pearl R. Wildlife Mgmt. Area, Honey Island Swamp Nature Tr., 31 August 1986, Steve Harsch 304 (RET 092-9); same location and date, Steve Harsch 305 (RET 093-6). MASSACHUSETTS: Middlesex County, Groton Conservation Area, 19 August 1989, Yala s.n. [Tulloss 8-19-89-B] (RET 245-4). RHODE ISLAND: Washington County, Carolina Management Area, 5 August 1988, Dobay 8-5-88-A (RET 239-8).

Commentary: Amanita brunneomaculata is characterized by its whitish to brownish pileus lacking striations on the margin, the floccose to patchy volval remnants on the pileus, and ellipsoid to elongate basidiospores (10.0–13.0 × 6.5–8.0 μm, Qm = 1.65 ± 0.19). This taxon is often in association with trees of the family Pinaceae.

Phylogenetically, A. brunneomaculata is sister to A. lanigera Yang-Yang Cui et al. (Fig. 1b). For comparisons between the two species see the commentary on A. lanigera. Amanita clarisquamosa, A. volvata (Peck) Lloyd, and A. rufobrunnescens are also morphologically similar to A. brunneomaculata. However, A. clarisquamosa has relatively narrower basidiospores (10.0–13.0 × 5.5–7.0 μm, Qm = 1.81 ± 0.19) (Imai 1933, 1938; Yang 1997, 2000, 2005, 2015; Imazeki et al. 2011). Amanita volvata, from the USA, has a basidioma with an obvious reddish brown color change when bruised or injured, a more floccose pileus, and a thicker subhymenium with 3–4 layers of inflated cells (Bas 1969; Jenkins 1978, 1986; Tulloss et al. 1995; this study). Furthermore, A. volvata is documented to be found in mixed forests in North American (Bas 1969; Jenkins 1978, 1986; Tulloss et al. 1995; this study). Amanita rufobrunnescens has narrower basidiospores (10.0–12.0 × 5.5–6.5 μm, Qm = 1.78 ± 0.17) and is dominant in subtropical forests with broad-leaved trees (Deng et al. 2016).

80. Amanita clarisquamosa (S. Imai) S. Imai apud E. J. Gilbert, Bres. Icon. Mycol. 27, Suppl. 1(2): 294 (1941).

Basionym: Amanitopsis clarisquamosa S. Imai, Bot. Mag. (Tokyo) 47: 430 (1933).

Pileus 4–10 cm in diam., dirty white (2B1), often yellow-brown (2B3–5) at center; volval remnants on pileus patchy or fibrous, gray-brown (1C2–4), brownish (4B2–4) to brown (4C2–4); margin shortly striate (0.1–0.2 R), sometimes appendiculate. Lamellae white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate. Stipe 6–13 cm long × 1–2 cm diam., white (1A1), with upper part covered with bran-shaped to floccose, gray-brown (1C2–4) squamules; basal bulb absent; volval remnants on stipe base saccate, 4–6 cm long × 2–4 (–5) cm diam., fleshy, outer surface white (1A1) to dirty white (2B1), inner surface dirty white (2B1). Annulus fragile and retained as pulverulence, white (1A1) to gray-brown (1C2–4) squamules on upper part of stipe.

Basidia 40–60 × 10–12 (–14) μm, clavate, 4-spored. Basidiospores [40/2/2] (9.5–) 10.0–13.0 × 5.5–7.0 μm, Q = (1.54–) 1.62–2.0 (–2.17), Qm = 1.81 ± 0.19, mostly elongate, sometimes ellipsoid or cylindrical, amyloid. Volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells fairly scarce to fairly abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in forests with Quercus and Abies; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study), India (Bhatt et al. 2003), Japan (Imai 1933, 1938; Imazeki et al. 2011) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. YUNNAN PROVINCE: Lijiang, Sandaowan, in a mixed forest with Quercus and Abies, altitude 3200 m, 1 August 1995, Zhu L. Yang 2116 (HKAS 29514). JAPAN. HOKKAIDO: Ishikari, Nopporo, altitude unknown, 11 October 1931, S. Imai (SAPA).

Commentary: Imai (1933, 1938) did not designate a type of A. clarisquamosa. Neither did Gilbert (1940) formally designate a type, but he illustrated five basidiospores of this species based on the collection made by S. Imai in the Nopporo forests on October 11, 1931. We examined two fragments of this collection but, unfortunately, no basidiospores were observed. In our multi-locus phylogenetic analysis (Fig. 1b), A. clarisquamosa is found to be related to A. volvata. However, collections of A. volvata listed under A. brunneomaculata have basidioma with an obvious reddish brown color change when bruised or injured, and is documented as being found in mixed forests in North American (Bas 1969; Jenkins 1978, 1986; Tulloss et al. 1995; this study). For detailed descriptions, comparisons with similar species, line drawings and images of A. clarisquamosa see Yang (1997, 2005, 2015).

81. Amanita lanigera Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 30d–f, 32.
Fig. 32

Microscopic features of Amanita lanigera (TYPE, HKAS 89030). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base, right side shows outer surface of volval remnants on stipe base. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825036

Etymology: lanigera, from laniger = fleecy, referring to its white basidioma densely covered with fleecy squamules of this species.

Type: CHINA. YUNNAN PROVINCE: Menghai County, in a forest with Castanopsis, Lithocarpus and Pinus, altitude 1200 m, 9 July 2014, Gang Wu 1293 (TYPE, HKAS 89030, GenBank Acc. No.: ITS = MH508420, nrLSU = MH486621, tef1-α  = MH508880, rpb2 = MH486074).

Basidioma (Fig. 30d–f) medium-sized. Pileus 6–9 cm in diam., plano-convex to applanate, lacking an umbo or depression at center, white (1A1) with brownish (4B3–5) tinge; volval remnants on pileus floccose to pulverulent, white (1A1), brownish (4B3–5) to brown (4C3–5), densely arranged over disk; margin non- or slightly striate when old, appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white to off-white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate, plentiful. Stipe 11–14 cm long × 1–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) with brown (4C3–5) tinge, covered with floccose, white (1A1), brownish (4B3–5) to brown (4C3–5) squamules; context white (1A1), unchanging, soft to hollow in center; basal bulb absent; volva saccate, 3–4 cm long × 4–6 cm diam., membranous, outer surface white (1A1) to brownish (4B3–5), inner surface brownish (4B3–5) to brown (4C3–5), 3–5 mm thick. Annulus present, pendant from attachment 3–4 cm below apex of stipe, white to off-white (1A1), easily broken and fugacious. Odor indistinct.

Lamellar trama bilateral. Mediostratum 40–70 μm wide, composed of abundant subfusiform, ellipsoid to elongate inflated cells (50–160 × 13–42 μm); filamentous hyphae abundant, 2–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subfusiform to ellipsoid inflated cells (25–75 × 10–30 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 32a) 30–50 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 12–25 × 8–18 μm. Basidia (Fig. 32a) 38–50 × 10–13 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 32b) [80/5/5] (9.0–) 10.0–12.0 (–14.0) × (6.0–) 7.0–8.5 (–9.0) μm, Q = (1.13–) 1.25–1.72 (–1.87), Qm = 1.49 ± 0.13, mostly ellipsoid, sometimes elongate, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (15–35 × 10–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 75–230 μm thick; upper layer (25–100 μm thick) gelatinized, composed of subradially to somewhat interwoven, thin-walled, colorless or yellowish, filamentous hyphae 2–8 μm wide; lower layer (50–130 μm thick) composed of radially and compactly arranged, filamentous hyphae 3–10 μm wide, colorless or yellowish; vascular hyphae scarce. Volval remnants on pileus composed of radially arranged elements: filamentous hyphae abundant to very abundant, 2–5 μm wide, yellowish to yellow-brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, ellipsoid, elongate to clavate, 50–100 × 12–25 μm, yellowish to yellow-brown, thin-walled, terminal or in chains of 2–3; vascular hyphae abundant. Interior of volval remnants on stipe base (Fig. 32c) composed of longitudinally arranged elements: filamentous hyphae very abundant, 3–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scattered to very abundant, subglobose, fusiform to ellipsoid or pyriform, 20–55 × 12–20 μm, colorless, thin-walled, terminal; vascular hyphae scarce. Outer surface of volval remnants on base (Fig. 32c) dominantly composed of very abundant filamentous hyphae, mixed with scarce pyriform inflated cells; inner surface gelatinized, similar to structure of interior part. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 80–520 × 15–45 μm; filamentous hyphae scattered to abundant, 2–8 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lancang County, in a forest dominated by Pinus, altitude 1300 m, 21 August 2016, LC-LJW 57 (HKAS 97561); same location, in a forest dominated by Pinus, altitude 2240 m, 15 August 2011, Gang Wu 704 (HKAS 75018); same location, in a forest with Pinus kesiya and Fagaceae, altitude 1160 m, 1 September 2017, Zhu L. Yang 6060 (HKAS 101249); Yongping County, in a mixed forest with Fagaceae and Pinaceae, altitude 2100 m, 31 July 2009, Gang Wu 67 (HKAS 57599).

Commentary: Amanita lanigera is characterized by its medium-sized basidioma, a white pileus lacking striations on the margin, floccose to pulverulent, white to brown volval remnants on the pileus and ellipsoid basidiospores (10.0–12.0 × 7.0–8.5 μm, Qm = 1.49 ± 0.17).

Phylogenetically, A. lanigera is sister to A. brunneomaculata (Fig. 1b). Indeed, they are also similar to each other in appearance. However, the latter differs by its indistinctly appendiculate pileus, relatively narrower basidiospores (10.0–13.0 × 6.5–8.0 μm, Qm = 1.65 ± 0.19) and its association with pine trees.

Amanita clarisquamosa and A. rufobrunnescens can be confused with A. lanigera due to their similar appearance. However, they have narrower basidiospores (10.0–13.0 × 5.5–7.0 μm, Qm = 1.81 ± 0.19 for A. clarisquamosa; 10.0–12.0 × 5.5–6.5 μm, Qm = 1.78 ± 0.17 for A. rufobrunnescens) (Imai 1933, 1938; Yang 1997, 2000, 2005, 2015; Imazeki et al. 2011; Deng et al. 2016). Furthermore, A. rufobrunnescens is found in subtropical forests with broad-leaved trees (Deng et al. 2016).

Amanita peckiana Kauffman and A. volvata, both described from the USA, are also similar to A. lanigera. However, A. peckiana has pinkish to pinkish cream volval remnants on the pileus and larger basidiospores (12.5–14.8 × 4.9–5.9 μm) (Peck 1913; Jenkins 1978, 1986; Tulloss et al. 1995). Collections of A. volvata, which are listed in the specimens examined under A. brunneomaculata, possess basidiomata with an obvious reddish brown color change when bruised or injured and relatively narrower basidiospores measuring [85/4/4] (9.0–) 10.0–12.5 (–13.0) × 6.0–7.5 (–9.0) μm, Q = (1.3–) 1.5–1.85 (–1.95), Qm = 1.67 ± 0.11.

82. Amanita parvicurta Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 30g, h, 33.
Fig. 33

Microscopic features of Amanita parvicurta (HKAS 100497). a Hymenium and subhymenium; b basidiospores. Bars: ab = 10 μm

MycoBank: MB 825037

Etymology: parvicurta, from parus = small, and curtus = short, referring to its small basidioma and short striations on pileal margin of this species.

Type: CHINA. YUNNAN PROVINCE: Lancang County, in a forest with Pinus kesiya and Fagaceae, altitude 1500 m, 30 August 2017, Zhu L. Yang 6026 (TYPE, HKAS 101215, GenBank Acc. No.: ITS = MH508490, nrLSU = MH486745, tef1-α  = MH508978, rpb2 = MH486169).

Basidioma (Fig. 30g, h) small. Pileus 3–6 cm in diam., plano-convex to applanate, lacking an umbo or depression at center, white (1A1) to dirty white (2B1), often brownish (3B2–4, 4B2–3) at center; volval remnants on pileus floccose to patchy, white (1A1), dirty white (2B1) to brownish (3B2–4, 4B2–3), easily falling away; margin slightly striate (ca. 0.2 R), slightly appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white to off-white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate, plentiful. Stipe 6–8 cm long × 0.5–1 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with furfuraceous, white (1A1) squamules; context white (1A1), soft to hollow in center; basal bulb absent; volva saccate, 2–3 cm high × 1.5–2 cm wide, membranous, outer surface white (1A1), dirty white (2B1) to brownish (3B2–4, 4B2–3), inner surface dirty white (2B1), 1–2 mm thick. Annulus fugacious. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–60 μm wide, composed of abundant subglobose, ellipsoid to clavate inflated cells (45–80 × 15–40 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid, fusiform to clavate inflated cells (30–70 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–7 μm wide. Subhymenium (Fig. 33a) 30–50 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 15–35 × 9–15 μm. Basidia (Fig. 33a) 30–50 × 8–11 μm, clavate, 4-spored; sterigmata 3–6 μm long; basal septa lacking clamps. Basidiospores (Fig. 33b) [40/2/2] (8.0–) 9.0–11.0 × 5.0–6.0 μm, Q = (1.6–) 1.67–2.16, Qm = 1.92 ± 0.13, elongate, sometimes cylindrical, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (25–45 × 13–30 μm), single and terminal or in chains of 2–3, thin-walled, colorless to brownish; filamentous hyphae abundant, 2–9 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 100–200 μm thick; upper layer (50–100 μm thick) gelatinized, composed of subradially to somewhat interwoven, thin-walled, colorless to brownish, filamentous hyphae 2–8 μm wide; lower layer (40–100 μm thick) composed of radially and compactly arranged, filamentous hyphae 4–10 μm wide, colorless to brownish; vascular hyphae scarce. Volval remnants on pileus composed of radially arranged elements: filamentous hyphae very abundant to nearly dominant, 3–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant, subglobose, fusiform to ellipsoid, 30–120 × 20–70 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base composed of two parts intergrading to each other. Outer part composed of longitudinally arranged elements: filamentous hyphae very abundant, 3–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce, but becoming fairly abundant toward inner part, subglobose, fusiform to ellipsoid, 30–80 × 20–60 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Inner part composed of longitudinally arranged elements: filamentous hyphae abundant, 3–10 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, fusiform to ellipsoid, 30–90 × 20–55 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volva dominantly composed of very abundant filamentous hyphae, 2–7 μm wide, occasionally mixed with scarce inflated cells; inner surface gelatinized, similar to structure of inner part, but with more abundant filamentous hyphae. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 50–380 × 20–45 μm; filamentous hyphae scattered to abundant, 2–10 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China.

Additional specimen examined: CHINA. FUJIAN PROVINCE: Shibazhai, in subtropical forest with Fagaceae, altitude 500 m, 4 September 2011, Xiang-Hua Wang 3091 (HKAS 100497).

Commentary: Amanita parvicurta is well distinguished by its small basidioma with a white pileus covered with floccose to patchy, white to brownish volval remnants, a white stipe decorated with concolorous squamules, elongate basidiospores (9.0–11.0 × 5.0–6.0 μm), and its occurrence in subtropical forests.

Phylogenetically, A. parvicurta is related to A. avellaneosquamosa (Fig. 1b). However, the latter differs in its relatively larger basidioma and longer striations (0.3–0.5 R) on the pileal margin (Imai 1933; Gilbert 1940, 1941a; Yang 1997, 2005, 2015; Yang and Doi 1999).

Amanita rufobrunnescens, a species described from Guangdong, China, can be confused with A. parvicurta. However, A. rufobrunnescens has a relatively larger basidioma which becomes reddish brown when bruised, and slightly broader basidiospores (10.0–12.0 × 5.5–6.5 µm, Qm = 1.78 ± 0.17) (Deng et al. 2016).

83. Amanita pinophila Yang-Yang Cui, Qing Cai & Zhu L. Yang, sp. nov.

Figures 30i–l, 34.
Fig. 34

Microscopic features of Amanita pinophila. a Hymenium and subhymenium (TYPE, HKAS 70167); b basidiospores (TYPE, HKAS 70167); c longitudinal section of outer part of volval remnants on stipe base, left side shows outer surface of volval remnants on stipe base (HKAS 70163); d longitudinal section of inner part of volval remnants on stipe base, right side shows inner surface of volval remnants on stipe base (HKAS 70163). Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825039

Etymology: pinophila, from pineus = piny, and -philus = -loving, referring to its basidioma associated with Pinus.

Type: CHINA. YUNNAN PROVINCE: Ninglang County, in a forest of Pinus yunnanensis, altitude 2300 m, 6 August 2011, Qing Cai 571 (TYPE, HKAS 70167, GenBank Acc. No.: ITS = MH508504, nrLSU = MH486759, tef1-α  = MH508986, rpb2 = MH486178, β-tubulin  = MH485682).

Basidioma (Fig. 30i–l) small to medium-sized. Pileus 3.5–6.5 cm in diam., plano-convex to applanate, lacking an umbo or depression at center, white (1A1), brownish (2B2–4) to brown (4B2–4); volval remnants on pileus floccose, concolorous with the pileus; margin slightly striate (0.1–0.2 R), non- or slightly appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate, plentiful. Stipe 4.5–7 cm long × 0.4–1.5 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1), brownish (2B2–4) to brown (4B2–4), covered with floccose to fibrous, concolorous squamules; context white (1A1), soft to hollow in center; basal bulb absent; volva saccate, 2–3 cm long × 2–4 cm diam., membranous, outer surface white (1A1), brownish (2B2–4) to brown (4B2–4), inner surface brownish (2B2–4) to brown (4B2–4), 1–3 mm thick. Annulus present, pendant from attachment 2–4 cm below apex of stipe, floccose, white (1A1), fragile. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–65 μm wide, composed of abundant subfusiform, ellipsoid to elongate inflated cells (35–100 × 15–35 μm); filamentous hyphae abundant, 2–10 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subfusiform to ellipsoid inflated cells (20–45 × 10–20 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 3–6 μm wide. Subhymenium (Fig. 34a) 30–60 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 8–15 × 5–15 μm. Basidia (Fig. 34a) 40–60 × 9–13 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 34b) [400/10/9] (8.0–) 10.0–12.0 × (5.0–) 5.5–7.0 μm, Q = (1.38–) 1.6–2.0 (–2.2), Qm = 1.81 ± 0.14, elongate, rarely ellipsoid or cylindrical, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (15–70 × 15–35 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–10 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 130–200 μm thick; upper layer (55–100 μm thick) gelatinized, composed of subradially to somewhat interwoven, thin-walled, colorless to yellowish, filamentous hyphae 2–6 μm wide; lower layer (75–100 μm thick) composed of radially and compactly arranged, filamentous hyphae 2–10 μm wide, colorless to yellowish; vascular hyphae scarce. Volval remnants on pileus dominantly composed of two parts intergrading to each other. Outer part of volval remnants on pileus composed of radially arranged elements: filamentous hyphae very abundant to nearly dominant, 2–8 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce, ellipsoid to clavate, 70–100 × 10–20 μm, colorless, thin-walled, often terminal; vascular hyphae scarce. Inner part of volval remnants on pileus composed of radially arranged elements: filamentous hyphae very abundant, 2–15 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, ellipsoid to clavate, 45–200 × 30–60 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Interior of volval remnants on stipe base (Fig. 34c, d) composed of longitudinally arranged elements: filamentous hyphae very abundant, 3–15 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells fairly abundant to abundant, subglobose, fusiform to ellipsoid, 40–110 × 20–80 μm, colorless, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Outer surface of volval remnants on stipe base (Fig. 34c) dominantly composed of very abundant filamentous hyphae, mixed with scarce, ellipsoid, fusiform to clavate, inflated cells; inner surface of volval remnants on stipe base (Fig. 34d) slightly gelatinized, similar to structure of interior part. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 50–200 × 25–60 μm; filamentous hyphae scattered to abundant, 4–13 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habit: Solitary to scattered under pine trees (e.g. Pinus yunnanensis); basidioma occurring in summer and autumn.

Known distribution: presently known from southwestern China.

Additional specimens examined: CHINA. YUNNAN PROVINCE: Lanping County, in a forest of Pinus yunnanensis, altitude 2350 m, 16 August 2011, Yan-Jia Hao 553 (HKAS 71662); Ninglang County, in a forest with Pinus, altitude 2440 m, 13 August 2010, Xue-Tai Zhu 131 (HKAS 68307); same location, in a forest dominated by Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Li-Ping Tang 1474 (HKAS 69935); same location, in a forest of Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Qing Cai 567 (HKAS 70163); same location, in a forest of Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Qing Cai 568 (HKAS 70164); same location, in a forest of Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Qing Cai 569 (HKAS 70165); same location, in a forest of Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Qing Cai 570 (HKAS 70166); same location, in a forest of Pinus yunnanensis, altitude ca. 2300 m, 6 August 2011, Qing Cai 572 (HKAS 70168).

Commentary: Amanita pinophila is characterized by its white to brownish pileus with striations on the margin, floccose volval remnants on the pileus, and mostly elongate basidiospores (10.0–12.0 × 5.5–7.0 μm, Qm = 1.81 ± 0.14). It is putatively associated with pine trees (e.g. Pinus yunnanensis).

Amanita pinophila formed a distinct lineage in our phylogenetic study (Fig. 1b). Amanita pinophila can be easily confused morphologically with A. volvata. However, specimens of A. volvata, which are listed in specimens studied under A. brunneomaculata, have basidiomata with a distinct reddish brown color change when bruised or injured and slightly narrower basidiospores (10.0–12.5 × 6.0–7.5 μm, Q = 1.5–1.85, Qm = 1.67 ± 0.11) (Bas 1969; Jenkins 1978, 1986; Tulloss et al. 1995; this study).

The East Asian A. avellaneosquamosa and A. clarisquamosa are also similar to A. pinophila in their white to brownish basidiomata. However, A. avellaneosquamosa has longer striations on the pileal margin, and relatively shorter basidiospores (9.0–11.0 × 5.0–7.0 μm) (Imai 1933; Gilbert 1940, 1941a; Yang 1997, 2005, 2015; Yang and Doi 1999). Amanita clarisquamosa has a relatively larger basidioma and longer basidiospores (Imai 1933, 1938; Yang 1997, 2000, 2005, 2015; Imazeki et al. 2011).

Amanita brunneomaculata and A. rufobrunnescens share some characters with A. pinophila. However, they have larger basidiomata. Furthermore, A. brunneomaculata has broader basidiospores (10.0–13.0 × 6.5–8.0, Qm = 1.65 ± 0.19). Amanita rufobrunnescens has a white to brownish basidioma with a distinct reddish brown color change when bruised, and occurs in subtropical forests with broad-leaved trees (Deng et al. 2016).

84. Amanita rufobrunnescens W. Q. Deng & T. H. Li, Phytotaxa 243 (2): 148 (2016).

Pileus 4–10 cm in diam., white (1A1) to dirty white (2B1); volval remnants on pileus submembranous to fibrous, brownish orange (6C4–6) to brown (6E4–6); margin striate (0.15–0.25 R), sometimes appendiculate; trama white (1A1), becoming reddish (7A2–4), brown (6E4–6) to reddish brown (7C2–4) when injured. Lamellae white (1A1), becoming grayish (1B1), gray-brown (2C2–4), brownish (3B2–4, 4B2–3) to brown (4D3–6) when dried; lamellulae mostly truncate. Stipe 7–15 cm long × 0.7–2.2 cm diam., white (1A1) to dirty white (2B1), densely covered with white (1A1) squamules; context white (1A1), becoming reddish (7A2–4), brown (6E4–6) to reddish brown (7C2–4) when injured, hollow in center; basal bulb absent; volval remnants on stipe base saccate, 3.5–6 cm long × 2.5–5 cm diam., fleshy, outer surface grayish orange (6B3–4) or brownish (5B2–4), inner surface white (1A1). Annulus fragile and retained as minute squamules on upper part of stipe.

Basidia 35–50 × 9–11 μm, clavate, 4-spored. Basidiospores [100/5/2] (9.5–) 10.0–12.0 (–13.0) × (5.0–) 5.5–6.5 (–7.0) μm, Q = 1.5–2.0 (–2.2), Qm = 1.78 ± 0.17, mostly elongate, occasionally ellipsoid or cylindrical, amyloid. Volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant; inflated cells scattered to fairly abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical broad-leaved forests with Fagaceae (e.g. Castanopsis fissa); basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Deng et al. 2016; this study).

Specimens examined: CHINA. GUANGDONG: Guangzhou, Tianluhu, in a broad-leaved forest, altitude 226 m, 17 June 2014, Y. W. Xia and Z. P. Song (HOLOTYPE, GDGM 42374); same location, in a broad-leaved forest, altitude 200–300 m, 4 July 2014, W. Q. Deng and Z. P. Song (PARATYPE, GDGM 44638).

Commentary: Amanita rufobrunnescens was described from China by Deng et al. (2016). This species is characterized by its white basidioma with a reddish brown color change when bruised or injured, a shortly striate pileal margin, elongate basidiospores (10–12 × 5.5–6.5 μm, Q = 1.5–2, Qm = 1.78 ± 0.17), and an association with subtropical broad-leaved trees (Deng et al. 2016).

Our phylogenetic study did not reveal any species closely related to A. rufobrunnescens (Fig. 1b). The North American A. volvata is morphologically similar to A. rufobrunnescens in the white basidiomata changing to reddish brown when bruised. However, specimens of A. volvata from the USA, which are listed in specimens examined under A. brunneomaculata, have a thicker subhymenium with 3–4 layers of inflated cells and relatively broader basidiospores (10–12.5 × 6–7.5 μm, Q = 1.5–1.85, Qm = 1.67 ± 0.11) (Bas 1969; Jenkins 1978, 1986; Tulloss et al. 1995; this study).

In China, A. brunneomaculata, A. clarisquamosa, A. lanigera and A. albidostipes may be confused with A. rufobrunnescens. However, these four species have basidiomata lacking the distinct reddish brown color change when bruised or injured (Imai 1933, 1938; Yang 1997, 2000, 2005, 2015; Imazeki et al. 2011; this study). In addition, A. brunneomaculata has broader basidiospores (10–13 × 6.5–8 μm, Q = 1.4–1.9, Qm = 1.65 ± 0.19) and is associated with pine trees. Amanita clarisquamosa is associated with Quercus and Abies. Amanita lanigera has broader basidiospores (10–12 × 7–8.5 μm, Q = 1.25–1.72, Qm = 1.49 ± 0.13) and is distributed in pine or mixed forests with Fagaceae and Pinaceae (Imai 1933, 1938; Yang 1997, 2000, 2005, 2015; Imazeki et al. 2011). Amanita pinophilia is associated with pine trees.

Amanita sect. Arenariae Zhu L. Yang, Yang-Yang Cui & Qing Cai, sect. nov.

MycoBank: MB 826870

Type: Amanita arenaria (O.K. Mill. & E. Horak) Justo, Mycologia 102(3): 682 (2010).

Diagnosis: Basidioma agaricoid or sequestrate; basidiospores thin-walled and smooth, amyloid, occasionally inamyloid; clamps absent in all parts of basidioma. When basidioma agaricoid, pileus covered with volval remnants; lamellulae truncate; stipe base turbinate. When basidioma sequestrate, gleba loculate; stipitate; basidia thick-walled. Known form south-western and western Australia on sandy soil and associated with Eucalyptus, Allocasuarina, Corymbia, Acacia, Daviesia, Eremophila, Melaleuca and Brachychiton.

Description: Basidioma agaricoid or sequestrate. When basidioma agaricoid, pileus applanate, pileal margin non-striate, appendiculate, context white to cream, becoming vinaceous-buff when bruised; volval remnants on pileus present; lamellae free, white to cream, lamellulae truncate; stipe subcylindric, stipe base turbinate; volval remnants on stipe base indistinct, as soft warts or small, marfinate rim; annulus present; basidia thin-walled; basidiospores amyloid; clamps absent in all parts of basidioma. When basidioma sequestrate, basidioma hypogeous to emergent, sequestrate and stipitate; pileus subglobose, oval or pulvinate, pileal margin enrolled and in contact with stipe, without exposing hymenium, context white, unchanging or becoming yellow when injured; volval remnants on pileus usually absent; gleba beige to brownish, loculate; stipe subcylindric and tapering downward, with its apex not extending to top of gleba; volval remnants on stipe base shortly limbate; annulus absent; basidia thick-walled when mature; basidiospores statismosporic, thin-walled, smooth; clamps absent in all parts of basidioma. On sandy soil, only known in south-western and western Australia and putatively associated with ectomycorhizal partners of Eucalyptus, Allocasuarina, Corymbia, Acacia, Daviesia, Eremophila, Melaleuca and Brachychiton.

Commentary: Previous data and our multi-locus phylogenetic study indicate that eight sequestrate species are currently known in Amanita (Malençon 1955; Miller and Horak 1992; Bougher 1999; Bougher and Lebel 2002; Hallen et al. 2004; Justo et al. 2010; Truong et al. 2017). Of these, A. arenaria was described as belonging to Torrendia and was then transferred to Amanita by Justo et al. (2010). It clustered with two agaricoid species, A. lesueurii E. M. Davison and A. wadjukiorum E. M. Davison, originally described from Western Australia and assigned to the stirps Strobiliformis of A. sect. Lepidella sensu Bas (Davison et al. 2013), as a distinct clade with strong statistic support. Furthermore, species in this clade are characterized by their agaricoid or sequestrate basidiomata, smooth basidiospores; the absence of clamps and their distributions in south-western and western Australia in association with Eucalyptus, Allocasuarina, Corymbia, Acacia, Daviesia, Eremophila, Melaleuca and Brachychiton. We therefore treated this clade as a novel section.

Our multigene-phylogenetic data (Fig. 1a) strongly indicate that A. sect. Arenariae shares a common ancestor with A. sect. Phalloideae, sect. Validae and sect. Strobiliformes. Fungi from these sections usually have amyloid basidiospores. However, this is not the case for the species A. arenaria. Justo et al. (2010) suggested that A. arenaria probably evolved from an amyloid-spored ancestor but this character was lost during the gasteromycetation process. To date, no species of A. sect. Arenariae has been reported from China.

Amanita sect. Phalloideae (Fr.) Quél., Mém. Soc. Emul. Montbéliard, Ser. II, 5: 66 (1872).

Basionym: Amanita [sect.] Phalloideae Fr., Monogr. Amanit. Sueciae: 3 (1854).

Lectotype: Amanita phalloides (Fr.: Fr.) Link., Handbuch zur Erkennung der Nutzbarsten und am häufigsten vorkommenden Gewächse: 272 (1833).

Diagnosis: Basidioma agaricoid; pileal surface glabrous or covered with membranous to pulverulent volval remnants; margin non-striate, non-appendiculate or rarely appendiculate; annulus membranous, persistent or sometimes fugacious; stipe with bulbous base; volva limbate and attached to bulb or breaking up as small warts; basidiospores amyloid; clamps absent.

Commentary: Our multi-locus phylogenetic evidence (Fig. 1b) indicates that the species in Amanita sect. Phalloideae could be separated into three subclades with high support (lethal Amanita subclade, A. hesleri-A. zangii subclade, and A. pseudogemmata-A. ballerina subclade), which may be treated as separate subsections. However, we will not formally propose such taxonomic changes until more taxa are studied. To date, 15 species of A. sect. Phalloideae have been reported from China, including one novel species and 14 known ones. Amanita subjunquillea var. alba Zhu L. Yang and A. areolata Oda et al. are treated as novel synonyms of A. subjunquillea S. Imai and A. zangii Zhu L. Yang et al., separately.

The lethal Amanita subclade includes all the lethal Amanita, which contain cyclopetide toxins and can cause fatal poisoning (Cai et al. 2014; Chen 2014; Chen et al. 2014, 2016; Tang et al. 2016). The fatalities caused by lethal Amanita species are presented in Fig. 35. Species in this subclade are characterized by a non-striate pileus rarely covered membranous volval remnants, attenuate lamellulae, a basal bulb with a limbate volva and the absence of clamps. To date, 12 lethal Amanita species have been found in China (see below).
Fig. 35

Poisonings and deaths caused by lethal mushrooms in Amanita sect. Phalloideae from China during 1994–2016

Only two species, A. hesleri and A. zangii, were recognized from the Amanita hesleri Bas subclade, and both were previously assigned to A. sect. Lepidella sensu Bas (1969). Recent molecular phylogenetic evidence indicates that both should be in A. sect. Phalloideae (Cai et al. 2014; Figs. 1b, S11), although neither amatoxins nor phallotoxins were detected in the basidiomata of A. zangii (Cai et al. 2014). Species in this subclade are characterized by their subconical to flat verrucose, felty to subfibrillose volval remnants on the pileus, appendiculate pileal margins, slenderly clavate to subfusiform base covered with floccose or warted volval remnants and the absence of clamps. To date, only A. zangii has been found in China (see below).

Currently, four species are known from the A. pseudogemmata subclade: A. pseudogemmata Hongo, A. levistriata D. T. Jenkins, A. ballerina Raspé et al. and A. franzii Zhu L. Yang et al. (Figs. 1b, S11). Species in this subclade are characterized by their striate pileal margins, truncate to nearly truncate lamellulae, marginate to sublimbate bulbs, (weakly) amyloid basidiospores and the absence of clamps. Amatoxins or phallotoxins have not been detected in A. pseudogemmata, A. ballerina and A. franzii (Cai et al. 2014; Thongbai et al. 2017). To date, A. pseudogemmata and A. franzii have been found in China (see below).

Key to the species of Amanita sect. Phalloideae from China

  1. 1.

    Pileal margin non-appendiculate; basal bulb subglobose or marginate; volval remnants on stipe base shortly limbate or limbate……….2

     
  2. 1.

    Pileal margin appendiculate; basal bulb narrowly fusiform to subclavate; volval remnants on stipe base non-limbate, pulverulent, floccose or warted………A. zangii

     
  3. 2.

    Pileal margin striate; volval remnants on pileus present; lamellulae mostly truncate; basal bulb marginate; volval remnants on stipe base shortly limbate; basidiospores weakly amyloid; cyclopetide toxins absent……….3

     
  4. 2.

    Pileal margin non-striate; volval remnants on pileus usually absent; lamellulae mostly attenuate; basal bulb subglobose; volval remnants on stipe base limbate; basidiospores distinctly amyloid; cyclopetide toxins present……….4

     
  5. 3.

    Pileus dirty yellow; volval remnants on pileus verrucose to pulverulent, brown-yellow to olive-yellow; stipe cream to white covered with yellow squamules; annulus yellow………A. pseudogemmata

     
  6. 3.

    Pileus dirty white; volval remnants on pileus verrucose to pulverulent, gray-brown to brown-gray; stipe white to dirty white covered with gray-brown squamules; annulus white to grayish………A. franzii

     
  7. 4.

    Basidia 2-spored……….5

     
  8. 4.

    Basidia 4-spored……….7

     
  9. 5.

    Pileus white, center occasionally cream, yellow or pale brown; lamellae white to cream; annulus apical to subapical……….6

     
  10. 5.

    Pileus brown, gray-brown to dark gray, center dark brown; lamellae pinkish; annulus nearly median………A. griseorosea

     
  11. 6.

    Basidioma larger with pileus ca. 4–8 cm diam.; basidiospores relatively larger, 9.5–12.0 × 9.0–11.5 μm………A. exitialis

     
  12. 6.

    Basidioma smaller with pileus ca. 3 cm diam.; basidiospores relatively smaller, 7.5–9.5 × 7.0–9.0 μm………A. parviexitialis

     
  13. 7.

    Basidioma white……….8

     
  14. 7.

    Basidioma differently colored………..13

     
  15. 8.

    Stipe covered with indistinctly white squamules; basidiospores smaller with average diameter less than 9 μm……….9

     
  16. 8.

    Stipe covered with distinctly white squamules; basidiospores larger with average diameter more than 9 μm, 9.0–11.0 × 8.5–10.5 μm………A. virosa

     
  17. 9.

    Basidioma relatively smaller; pileus ca. 3–6 cm in diam., purely white and occasionally umbonate at center……….10

     
  18. 9.

    Basidioma relatively larger; pileus ca. 5–10 cm in diam., umbonate and pinkish at center, white at margin……….12

     
  19. 10.

    Pileipellis composed of abundant long ellipsoidal to clavate inflated cells……….11

     
  20. 10.

    Pileipellis composed of radially arranged filamentous hyphae………A. subjunquillea (albino form)

     
  21. 11.

    Interior of volval limb composed of very abundant filamentous hyphae and evenly distributed scattered inflated cells; basidioma distributed in tropical to subtropical China………A. rimosa

     
  22. 11.

    Interior of volval limb composed of two layers intergrading into each other, the inner layer composed of abundant filamentous hyphae and abundant inflated cells, outer layer composed of dominantly filamentous hyphae; basidioma distributed in subtropical to temperate China………A. molliuscula

     
  23. 12.

    Stipe covered with finely fibrillose, evenly distributed squamules below annulus; basidia 30–45 × 9–11 μm; basidiospores 6.0–8.0 × 6.0–7.5 μm………A. pallidorosea

     
  24. 12.

    Stipe covered with distinctly lacerate, locally distributed squamules below annulus; basidia 35–50 × 11–13 μm; basidiospores 8.0–10.0 × 7.5–9.0 μm………A. subpallidorosea

     
  25. 13.

    Pileus brown, gray-brown to dark brown; stipe covered with dirty white, grayish or grayish brown squamules……….14

     
  26. 13.

    Pileus brownish yellow, dirty citrine-yellow to mustard yellow, occasionally white; stipe white to yellowish covered with yellowish fibrillose squamules………A. subjunquillea

     
  27. 14.

    Basidioma smaller with pileus 3–6 cm diam.; basal bulb on stipe globose to subglobose……….15

     
  28. 14.

    Basidioma larger with pileus 7–12 cm diam.; basal bulb on stipe subclavate to napiform………A. fuligineoides

     
  29. 15.

    Stipe darker, covered with brown to grayish brown squamules; basidiospores globose, rarely subglobose, 7.0–9.0 × 7.0–9.0 μm………A. subfuliginea

     
  30. 15.

    Stipe lighter, covered with white, gray to brownish squamules; basidiospores globose to subglobose, 7.0–9.0 × 6.5–8.5 μm………A. fuliginea

     

85. Amanita exitialis Zhu L. Yang & T. H. Li, Mycotaxon 78: 439 (2001).

Pileus 4–8 cm in diam., white (1A1), but often cream (1A2) at center; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 7–9 cm long × 0.5–1.5 (–2) cm diam., white (1A1), glabrous or covered with fibrous, white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1–2 (–3) cm diam.; volva limbate, membranous, with free limb up to 2 cm in height, both surfaces white (1A1). Annulus apical to subapical, white (1A1), persistent. All parts of basidioma becoming yellow immediately with 5% KOH solution.

Basidia 27–55 × 10–15 μm, clavate, 2-spored. Basidiospores [200/9/9] (9.0–) 9.5–12.0 (–14.5) × (8.5–) 9.0–11.5 (–13) μm, Q = 1.0–1.15 (–1.17), Qm = 1.08 ± 0.04, globose to subglobose, amyloid. Interior of volval remnants on stipe base composed of irregularly, compactly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in forests dominated by Fagaceae; basidioma occurring from early spring to summer.

Distribution: Known in southern and southwestern China (Yang and Li 2001; Yang 2005, 2015; Cai et al. 2016) and India (Bhatt et al. 2003).

Specimens examined: CHINA. GUANGDONG ROVINCE: Guangzhou, Baiyunshan, in a subtropical forest dominated by Fagaceae, altitude 110 m, date unknown, Ping Zhang A1 (HKAS 75774); Guangzhou, Fenghuangshan, in a subtropical forest dominated by Fagaceae, altitude 350 m, date unknown, Ping Zhang G10 (HKAS 75776); Guangzhou, South China Agricultural University, in a subtropical forest dominated by Fagaceae, altitude 34 m, date unknown, Ping Zhang C19 (HKAS 75775); Guangzhou, Tianluhu, in a subtropical broad-leaved forest with Fagaceae, altitude 190 m, 25 March 2001, Zuo-Hong Chen s.n. (HKAS 38162); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 50 m, 23 March 2000, Tai-Hui Li s.n. (TYPE, GDGM 20241). YUNNAN ROVINCE: Nanjian County, Gonglang, in a subtropical broad-leaved forest with Fagaceae, altitude 1570 m, 2 August 2015, Qing Cai 1358 (HKAS 90175); Nanjian County, Lingbaoshan, in a subtropical forest with Lithocarpus and Castanopsis, altitude 2350 m, 3 August 2014, Qing Cai 1207 (HKAS 83674); Kunming, Qiongzhusi, in a subtropical broad-leaved forest with Quercus, altitude 2200 m, 5 October 2011, Bang Feng 1192 (HKAS 74673); Puer, in a subtropical broad-leaved forest with Fagaceae, altitude 1400 m, 28 June 2014, Kuan Zhao 432 (HKAS 89095).

Specimens of A. suballiacea examined: USA. CONNECTICUT: Middlesex County, East Haddam, Devil’s Hopyard State Park, 4 September 2011, Kevin Scalzo s.n. [Tulloss 9-4-11-K] (RET 490-1). MICHIGAN: McComb County, Shelby Township, ca. Washington, Stony Brook Metropolitan Park [42.73°N/83.07°W, 250–260 m], 30–31 August 2011, Sandy Sheine s.n. (RET 491-7). PENNSYLVANIA: Clarion County, Clarion, State Game Lands 63, Robert’s Hill Road [41.1421°N/79.2830°W, 419 m], 6 August 2011, Bill Bynum s.n. [Tulloss 8-6-11-C] (RET 478-6).

Commentary: Amanita exitialis was described from China by Yang and Li (2001), and subsequently reported from India (Bhatt et al. 2003). This species belongs to the lethal Amanita subclade in A. sect. Phalloideae. Cyclopetide toxins, in particular α-amanitin and β-amanitin, are detected in all parts of the basidioma and at different stages of development (Deng et al. 2011; Xue et al. 2011; Hu et al. 2012; Li et al. 2014b; Chen et al. 2016; Tang et al. 2016). It is the leading cause of mushroom poisonings in Guangdong Province, China, where from 2000 to 2014, 22 cases of mushroom poisoning were reported, resulting in 89 affected individuals and 45 deaths (Yang and Li 2001; Deng et al. 2011; Xue et al. 2011; Hu et al. 2012; Cai et al. 2014; Chen 2014; Chen et al. 2014, 2016; Li et al. 2014b; Tang et al. 2016).

Our multi-locus phylogenetic analysis (Fig. 1b) shows that A. exitialis is sister to the North American A. suballiacea (Murrill) Murrill (Cai et al. 2014). Examination of specimens of A. suballiacea from North America indicates that it has 4-spored basidia and smaller, broadly ellipsoid to ellipsoid basidiospores (8.5–10.5 × 6.5–8.5 μm, Q = 1.2–1.4, Qm = 1.29 ± 0.08) (Murrill 1941; Jenkins 1979, 1986; this study). Furthermore, the outer layer of volval remnants on the stipe base in A. suballiacea is composed of wide filamentous hyphae up to 20 μm in diam., while those in A. exitialis are 2–7 μm in diam. For detailed descriptions, comparisons with similar species and images of A. exitialis see Yang and Li (2001), Yang (2005, 2015) and Cai et al. (2016).

86. Amanita franzii Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 30m–o, 36.
Fig. 36

Features of Amanita franzii. a Basidioma (HKAS 91231); b Hymenium and subhymenium (TYPE, HKAS 79559); c Basidiospores (TYPE, HKAS 79559). Bars: a = 2 cm, bc = 10 μm

MycoBank: MB 825038

Etymology: The epithet “franzii” is in memory of the late Prof. Dr. Franz Oberwinkler (1939–2018).

Type: CHINA. YUNNAN PROVINCE: Kunming, Qiongzhusi, in a mixed forest with Fagaceae and Pinaceae, altitude 2000 m, 6 September 2012, Qing Cai 880 (TYPE, HKAS 79559, GenBank Acc. No.: nrLSU = MH486523, tef1-α  = MH508799, rpb2 = MH485992, β-tubulin  = MH485514).

Basidioma (Figs. 30m–o, 36a) medium-sized. Pileus 5–9 cm diam., convex to applanate, dirty white (2B1) to pale yellow-brown (1B2–3), becoming paler towards margin; volval remnants on pileus verrucose, bran-like to pulverulent, sometimes felted, gray-brown (2D2–4) to brown-gray (2C2–4); margin slightly striate (0.1–0.2 R), non-appendiculate; trama dirty white (2B1), unchanging. Lamellae free, crowded, white (1A1) to cream (1A2); lamellulae truncate to nearly truncate (Fig. 36a), plentiful. Stipe 7–10 cm long × 0.5–1.8 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) to dirty white (2B1), covered with gray-brown (2D2–4) squamules; context white (1A1), unchanging; basal bulb marginate, 1.5–3 cm diam., dirty white (2B1) to grayish (1B1), with upper edge shortly limbate, dirty white (2B1) to grayish (1B1) volval remnants. Annulus present, superior, membranous, white (1A1) at upper surface, grayish (1B1) at lower surface. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–50 μm wide, composed of abundant ellipsoid to clavate inflated cells (50–200 × 20–35 μm); filamentous hyphae abundant, 5–12 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant ellipsoid to clavate inflated cells (25–75 × 13–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–8 μm wide. Subhymenium (Fig. 36b) 40–60 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregular cells, 10–20 × 8–15 μm. Basidia (Fig. 36b) 45–60 × 10–12 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 36c) [105/6/5] 8.5–10.5 (–11.5) × 6.5–7.5 (–8.5) μm, Q = 1.12–1.46, Qm = 1.34 ± 0.11, broadly ellipsoid to ellipsoid, rarely subglobose, weakly amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of globose, subglobose, fusiform to elongate or sphaeropedunculate inflated cells (20–40 × 15–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–7 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 125–150 μm thick; upper layer (30–60 μm thick) non- or slightly gelatinized, composed of subradially to somewhat interwoven, thin-walled, colorless, filamentous hyphae 2–8 μm wide; lower layer (60–75 μm thick) composed of radially and compactly arranged, colorless, filamentous hyphae 4–15 μm wide; vascular hyphae scarce. Volval remnants on pileus composed of irregularly to subvertically arranged elements: filamentous hyphae fairly abundant, 2–18 μm wide, colorless, brown to gray-brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant, globose, subglobose to ellipsoid, 30–110 × 20–85 μm, colorless, brown to gray-brown, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 50–300 × 20–50 μm; filamentous hyphae scattered to abundant, 5–15 μm wide; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from central and southwestern China.

Additional specimens examined: CHINA. HUNAN ROVINCE: Yizhang County, Mangshan, in a subtropical forest, altitude 900 m, 3 July 1997, Zuo-Hong Chen 3678 (HKAS 37013). YUNNAN PROVINCE: Kunming, Yeyahu, in a mixed forest with Fagaceae and Pinaceae, altitude 2000 m, 10 August 2013, Yan-Jia Hao 1041 [HKAS 82832, as Amanita sp. 17 in Yang (2015)]; Kunming, Qiongzhusi, in a mixed forest with Fagaceae and Pinaceae, altitude 2000 m, 1 August 2015, Jing Li 203 (HKAS 91231); same city, Yeyahu, in a mixed forest with Fagaceae and Pinaceae, altitude 2000 m, 18 August 2012, Qing Cai 828 (HKAS 77321).

Commentary: Amanita franzii is characterized by the rice bran-like to pulverulent, gray-brown volval remnants on the pileus, a marginate basal bulb, weakly amyloid basidiospores and the absence of clamps.

Our molecular phylogenetic analysis (Fig. 1b) indicates that A. franzii clusters as a lineage in the A. pseudogemmata subclade of A. sect. Phalloideae and is closely related to A. ballerina. However, A. ballerina can be distinguished from A. franzii by its relatively smaller basidioma and a white pileus covered with white to cream patchy volval remnants (Thongbai et al. 2017).

Amanita pseudogemmata is comparable to A. franzii in the marginate basal bulb. However, A. pseudogemmata differs from A. franzii by its yellow pileus covered with yellow to yellow-brown volval remnants, and a yellow stipe with its basal bulb covered with collar-like, white to yellowish volval remnants (Hongo 1974b; Yang and Doi 1999; Yang 2005, 2015; this study).

The marginate bulb of Amanita franzii and the nature of the volval remnants are reminiscent of species of A. subsect. Limbatulae, including A. mutabilis Beardslee, A. parva (Murrill) Murrill, and A. praelongispora (Murrill) Murrill, which were described from the USA. However, these American taxa have sublimbate stipe bases and proportionally narrower basidiospores (Beardslee 1919; Murrill 1941, 1945; Bas 1969; Jenkins 1979, 1986; Tulloss 1984). In addition, the latter two species have clamps (Beardslee 1919; Murrill 1941, 1945; Bas 1969; Jenkins 1979, 1986; Tulloss 1984).

87. Amanita fuliginea Hongo, Journ. Jap. Bot. 28: 69 (1953).

Pileus 3–6 cm in diam., dark gray (1A1–3), fuliginous (3E1–4), dark brown (5E1–3) to black (1F1–4), often darker at center, with innate, dark fibrils; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 6–10 cm long × 0.5–1 cm diam., white (1A1) to grayish (1D1–4), often covered with minute, fibrous, brownish (3D2–5) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1–2.5 cm diam.; volva limbate, membranous, with free limb up to 3–8 cm in height, outer surface white (1A1) to dirty white (2B1), inner surface white (1A1). Annulus apical to subapical, gray (1E1–3) to dirty white (2B1), persistent.

Basidia 30–40 × 10–13 μm, clavate, 4-spored. Basidiospores [220/21/19] 7.0–9.0 (–11.0) × (6.0–) 6.5–8.5 (–10.0) μm, Q = 1.0–1.12 (–1.21), Qm = 1.06 ± 0.05, globose to subglobose, rarely broadly ellipsoid, amyloid. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring from late spring, summer to autumn.

Distribution: Known in eastern, southern, central and southwestern China (Yang 1997, 2005, 2015; Cai et al. 2016; this study), Japan (Hongo 1953; Imazeki et al. 2011) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 915 m, 24 July 2017, Yan-Jia Hao 1557 (HKAS 100628). FUJIAN PROVINCE: Sanming, in a subtropical forest, altitude 265 m, 27 August 2007, Ping Zhang 635 (HKAS 52707). GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 470 m, 11 May 2012, Fang Li 249 (HKAS 75781); same location, in a forest dominated by Fagaceae, altitude ca. 375 m, 17 May 2012, Fang Li 310 (HKAS 75780); same location, in a subtropical forest dominated by Fagaceae, altitude 150 m, 25 June 2012, Fang Li 519 (HKAS 78472); same location, in a subtropical forest dominated by Fagaceae, altitude 200 m, 26 June 2012, Fang Li 544 (HKAS 75782); same location, in a subtropical forest dominated by Fagaceae, altitude 500 m, 4 July 2012, Fang Li 603 (HKAS 78481); same location, in a subtropical forest dominated by Fagaceae, altitude 700 m, 5 July 2012, Fang Li 630 (HKAS 78488); Guangzhou, Baiyunshan, in a subtropical mixed forest with Castanopsis, Lithocarpus and Pinus, altitude 380 m, 2 September 1998, Zhu L. Yang 2495 (HKAS 32493). HUNAN PROVINCE: Changsha, Fulin, in a subtropical mixed forest with Lithocarpus and Pinus massoniana, altitude 400 m, 12 June 2001, Zhu L. Yang 3051 (HKAS 38129); same location, in a subtropical broad-leaved forest, altitude 150 m, 18 June 2000, Zuo-Hong Chen 4002 (HKAS 37009); Yizhang County, Mangshan, in a subtropical forest, altitude 940 m, collection date unknown, Ping Zhang 749 (HKAS 79685); Yizhang County, altitude 1000 m, 15 June 2001, Zhu L. Yang 3054 (HKAS 38132). JIANGXI PROVINCE: Fuzhou, Linggufeng, in a subtropical, broad-leaved forest with Fagaceae, altitude 40 m, 20 June 2012, Gang Wu 961 (HKAS 77132). SICHUAN PROVINCE: Emeishan, in a forest with Castanopsis and Quercus, altitude 1400 m, 1 October 1999, Zhu L. Yang 2753 (HKAS 34219). YUNNAN PROVINCE: Yiliang County, Xiaocaoba, altitude 1800 m, 17 September 1998, Zhu L. Yang 2500 (HKAS 32676); Xichou County, Fadou, in a forest with Castanopsis and Lithocarpus, altitude 1550 m, 18 June 1992, Zhu L. Yang 1746 (HKAS 32521). JAPAN. TOTTORI: Iwami-gun, Kokufu-cho, Okamasu, under Castanopsis cuspidate, altitude unknown, 11 July 1981, E. Nagassawa s.n. (HKAS 35322). OTSU: Mii-dera, in a forest of Castanopsis cuspidate, mixed with planted Chamaecyparis obtusa, altitude unknown, 4 August 1953, Hongo Herb. 711 (TNS-F-237540).

Commentary: Amanita fuliginea was described from Japan by Hongo (1953), and subsequently reported from China and Thailand (Yang 1997, 2005, 2015; Sanmee et al. 2008). This species belongs to the lethal Amanita subclade in A. sect. Phalloideae. It has been reported from East and southeast Asia (Hongo 1953; Yang 1997, 2005, 2015; Sanmee et al. 2008; Imazeki et al. 2011; Parnmen et al. 2016). Amanita fuliginea, containing high content of cyclopetide toxins, is a main cause of fatal mushroom poisoning in southern and central China, where during 1994–2012, 33 poisoning cases were reported, resulting in 352 affected individuals and 79 deaths (Chen et al. 2003, 2014, 2016; Parnmen et al. 2016; Tang et al. 2016; Zhou et al. 2017).

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. fuliginea is closely related to A. subfuliginea Qing Cai et al. For detailed descriptions, comparisons with similar species and images of A. fuliginea see Yang (1997, 2005, 2015) and Cai et al. (2016).

88. Amanita fuligineoides P. Zhang & Zhu L. Yang, Fungal Divers. 42: 122 (2010).

Pileus 7–14 cm in diam., gray-brown (2E2–5), fuliginous (3E1–4) to black (1F1–4), often darker at center, with innate, dark fibrils; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 10–14 (–20) cm long × 0.8–1.5 (–2) cm diam., white (1A1) to grayish (1D1–4), covered with minute, fibrous to snakeskin-shaped, gray-brown (4E2–5) squamules; context white (1A1), unchanging; basal bulb napiform to clavate, ca. 1–2.5 cm diam.; volva limbate, membranous, with free limb up to 1.5 cm in height, both surfaces white (1A1). Annulus apical to subapical, white (1A1) to grayish (1D1–4), persistent.

Basidia 30–45 × 9–12 μm, clavate, 4-spored. Basidiospores [80/5/5] (7.0–) 7.5–9.5 (–10.0) × 7.0–9.0 μm, Q = 1.0–1.13 (–1.21), Qm = 1.06 ± 0.05, globose to subglobose, amyloid. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved or mixed forests with Fagaceae and Pinaceae, basidioma occurring from summer to autumn.

Distribution: Known in central and southwestern China (Zhang et al. 2010; Yang 2015; Cai et al. 2016; this study), and Thailand (Thongbai et al. 2017).

Specimens examined: CHINA. HUNAN PROVINCE: Yizhang County, Mangshan, in a subtropical broad-leaved forest with Fagaceae, altitude 1100 m, 6 September 2007, Ping Zhang 664 (TYPE, HKAS 52316); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 900 m, 3 September 2007, Ping Zhang 647 (HKAS 52717); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 900 m, 5 September 2007, Ping Zhang 657 (HKAS 52727); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 800 m, 12 September 2005, Ping Zhang 510 (HKAS 49688). YUNNAN PROVINCE: Nanjian County, Lingbaoshan, in a subtropical forest with Castanopsis and Pinus, altitude 2350 m, 4 August 2014, Qing Cai 1225 (HKAS 83694).

Commentary: Amanita fuligineoides was described from China by Zhang et al. (2010), and subsequently reported from Thailand (Thongbai et al. 2017). This species is a member of the lethal Amanita subclade in A. sect. Phalloideae. Cyclopetide toxins, including α-amanitin, β-amanitin and phalloidin, have been detected in the basidioma of A. fuligineoides, thus, acute liver failure would be caused by ingestion of this species (Chen et al. 2016; Tang et al. 2016; Thongbai et al. 2017).

Our phylogenetic analysis (Fig. 1b) indicates that A. fuligineoides is closely related to A. brunneitoxicaria Thongbai et al., which is in agreement with the conclusions of Thongbai et al. (2017). However, A. brunneitoxicaria differs from A. fuligineoides by its relatively smaller basidioma with pileus ca. 3.5–4.5 in diam., a paler colored stipe (white to pale grayish brown) and broadly ellipsoid to ellipsoid basidiospores (7.9–9.8 × 6.8–7.7 μm, Q = 1.2–1.42, Qm = 1.27 ± 0.3) (Thongbai et al. 2017). For detailed descriptions, comparisons with similar species and images of A. fuligineoides see Zhang et al. (2010), Yang (2015) and Cai et al. (2016).

89. Amanita griseorosea Qing Cai, Zhu L. Yang & Yang-Yang Cui, Mycologia 108 (5): 997 (2016).

Pileus 3–6 cm in diam., gray (4C2–4) to grayish brown (4D3–4), with innate, dark gray, radiating fibrils; volval remnants on pileus absent; margin non-striate, sometimes radially rimose, non-appendiculate; trama white, unchanging. Lamellae initially white (1A1), but becoming pinkish (7A2–3, 8A2) when mature; lamellulae attenuate. Stipe 5–7 cm long × 0.5–1 cm diam., white to dirty white, covered with minute, white to dirty white squamules; context white (1A1), unchanging; basal bulb subglobose, 1–1.5 cm diam.; volva limbate, membranous, with free limb up to 1 cm in height, both surfaces white (1A1). Annulus median, white (1A1), persistent.

Basidia 32–48 × 10–13 μm, clavate, 2-spored. Basidiospores [180/9/7] (7.5–) 8.5–10.5 (–11.5) × (7.0–) 8.0–10.0 μm, Q = 1.0–1.14 (–1.4), Qm = 1.06 ± 0.05, globose to subglobose, amyloid. Interior of volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary or gregarious in tropical to subtropical forests with Fagaceae; basidioma occurring in summer.

Distribution: Known from southern and southwestern China (Cai et al. 2016; this study).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a forest dominated by Fagaceae, altitude ca. 500 m, 29 April 2013, Fang Li 1278 (HKAS 79176). HAINAN PROVINCE: Baisha County, Yinggeling, in a tropical to subtropical forest, altitude 1300 m, 28 July 2009, Nian-Kai Zeng 385 (HKAS 77333); Lingshui County, Diaoluoshan, in a tropical to subtropical forest, altitude 900 m, 29 July 2010, Nian-Kai Zeng 757 (HKAS 77334); detailed location and altitude unknown, in a tropical to subtropical forest, 2 September 1960, Rong Liu 2313 (HMAS 32819); Jianfengling, in a tropical to subtropical forest, altitude 109 m, 29 September 1987, Tai-Hui Li s.n. (GDGM 12541); Qiongzhong County, Limushan, in a tropical to subtropical forest, altitude 1000 m, 22 July 2009, Nian-Kai Zeng 307 (TYPE, HKAS 77332). YUNNAN PROVINCE: Mengla County, Xishuangbanna, in a subtropical forest with Fagaceae, altitude 1040 m, 6 July 2014, Gang Wu 1267 [HKAS 83450, as Amanita sp. 18 in Yang (2015)].

Commentary: Amanita griseorosea corresponds to Amanita sp. 7 and Amanita sp. 18 in Cai et al. (2014) and Yang (2015), respectively. It is a member of the lethal Amanita subclade in A. sect. Phalloideae. β-amanitin has been detected in the basidiomata of A. griseorosea (Cai et al. 2014; Chen et al. 2016).

Phylogenetically, A. griseorosea is closely related to A. molliuscula Qing Cai et al. (Fig. 1b). However, A. molliuscula has a white basidioma, 4-spored basidia, smaller basidiospores (7.5–9.0 × 7.0–8.0 μm, Q = 1.0–1.14, Qm = 1.07 ± 0.06), and abundant inflated cells in the pileipellis and in the inner layer of the volval limb (Cai et al. 2016, this study). For detailed descriptions, comparisons with similar species and images of A. griseorosea see Yang (2015) and Cai et al. (2016).

90. Amanita molliuscula Qing Cai, Zhu L. Yang & Yang-Yang Cui, Mycologia 108 (5): 999 (2016).

Pileus 4–6 cm in diam., white (5A1), sometimes pale buff (5A2–3) at center; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white, unchanging. Lamellae white (11A1) to cream-colored (2A2); lamellulae attenuate. Stipe 8–10 cm long × 0.6–1 cm diam., white (1A1), covered with fibrous, white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1.5 cm diam.; volva limbate, somewhat soft membranous, both surfaces white (1A1). Annulus apical to subapical, white (1A1), persistent.

Basidia 30–45 × 8–12 μm, clavate, 4-spored. Basidiospores [100/3/3] (7.0–) 7.5–9.0 (–10.0) × (6.5–) 7.0–8.0 (–9.0) μm, Q = 1.0–1.14 (–1.25), Qm = 1.07 ± 0.06, globose to subglobose, amyloid. Pileipellis composed of two layers: upper layer slightly or non-gelatinized, composed of subradially arranged, ellipsoid to clavate inflated cells, mixed with filamentous hyphae; lower layer composed of radially and compactly arranged, filamentous hyphae. Interior of volval remnants on stipe base composed of two layers intergrading into each other: Outer layer dominantly composed of longitudinally arranged filamentous hyphae, mixed with scarce long clavate inflated hyphae, lacking any distinct globose to ellipsoidal inflated cells. Inner layer composed of longitudinally arranged elements: filamentous hyphae abundant; inflated cells abundant, subglobose, ellipsoid to long clavate. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in subtropical to temperate forests with Fagaceae and Pinaceae; basidioma occurring in summer to autumn.

Distribution: Known from central and northwestern China (Cai et al. 2016; this study).

Specimens examined: CHINA. SHAANXI PROVINCE: Zhouzhi County, Taibaishan, in a forest with Quercus and Pinus, altitude 1400 m, 15 July 2012, Xiang-Hua Wang 2916 (TYPE, HKAS 77324). HUBEI PROVINCE: Yichang, Shennongjia, in a forest with Quercus, altitude 1850 m, 15 July 2012, Qing Cai 801 (HKAS 75555). JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest, altitude 1000 m, 5 September 2008, collector unknown (HMJAU 20469).

Commentary: Amanita molliuscula corresponds to Amanita sp. 6 in Cai et al. (2014). It belongs to the lethal Amanita subclade of A. sect. Phalloideae. Cyclopetide toxins, including α-amanitin and β-amanitin, have been detected in basidiomata of A. molliuscula (Cai et al. 2014). Phylogenetically, A. molliuscula and A. griseorosea clustered together with strong statistic support (Fig. 1b). For comparisons between them see the commentary on A. griseorosea. For detailed descriptions, comparisons with similar species and images of A. molliuscula see Cai et al. (2016).

91. Amanita pallidorosea P. Zhang & Zhu L. Yang, Fungal Divers. 42: 125 (2010).

Pileus 5–10 cm in diam., often umbonate at center, pinkish (7A2–4) at center, white (1A1) at margin, sometimes pure white (1A1) over entire disk; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white, unchanging. Lamellae white (11A1); lamellulae attenuate. Stipe 8–15 cm long × 0.6–1.2 cm diam., white (1A1), dirty white (2B1) to pale yellow-brown (2B2–4), covered with fibrous, white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1.2–2.2 cm diam.; volva limbate, membranous, with free limb up to 2 cm in height, both surfaces white (1A1). Annulus superior, white (1A1), persistent. All parts of basidioma becoming yellow immediately with 5% KOH solution.

Basidia 30–45 × 9–11 μm, clavate, 4-spored. Basidiospores [189/22/19] 6.0–8.0 (–9.0) × (5.0–) 6.0–8.0 μm, Q = 1.0–1.14 (–1.25), Qm = 1.07 ± 0.06, globose to subglobose, rarely broadly ellipsoid, amyloid. Interior of volval remnants on stipe base composed of irregularly, compactly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical to temperate broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring from summer to autumn.

Distribution: Known in northeastern, eastern, northwestern, central and southwestern China (Zhang et al. 2010; Yang 2015; Cai et al. 2016; this study), India (Bhatt et al. 2017), Japan (this study), Pakistan (Kiran et al. 2018), Republic of Korea (Kim et al. 2013a) and Russian Far East (Malysheva et al. 2014).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 955 m, 23 July 2017, Yan-Jia Hao 1550 (HKAS 100625). GANSU ROVINCE: Tianshui, Dongcha, in a temperate forest with Quercus, altitude 1300 m, 25 July 2011, Xiao-Fei Shi 746 (HKAS 75786); Tianshui, Maijishan, in a temperate forest, altitude 1600 m, 4 August 2012, Qi Zhao 1626 (HKAS 78885). GUIZHOU PROVINCE: Daozhen County, Yangxi, in a subtropical forest with Castanopsis, Fagus and Pinus, altitude 900 m, 28 July 2010, Xiang-Hua Wang 2423 (HKAS 77327); Suiyang County, Kuankuoshui, in a subtropical forest with Castanopsis and Fagus, altitude 1500 m, 25 July 1010, Xiang-Hua Wang 2407 (HKAS 77329). HENAN PROVINCE: Neixiang County, Taohuayuan, altitude 500–700 m, 31 July 2010, Xiang-Hua Wang 2440-a (HKAS 75784). HUBEI PROVINCE: Fan County, Xihao, in a forest with Quercus, altitude 1400 m, 30 July 2010, Xiang-Hua Wang 2933 (HKAS 77337); Shiyan, in a mixed forest with Fagaceae and Pinaceae, altitude 1000 m, 3 July 2013, Yan-Jia Hao 934 (HKAS 80214); Xingshan County, Nanyang, in a forest with Quercus, altitude 1450 m, 11 July 2012, Qing Cai 728 (HKAS 75483); Yichang, Shennongjia, in a forest with Fagaceae, altitude 1000 m, 2 July 2013, Yan-Jia Hao 933 (HKAS 80213). JILIN PROVINCE: Huinan County, in a temperate forest, altitude 720 m, 29 August 2007, Ying-Hai Bao s.n. (HMJAU 5846). LIAONIANG PROVINCE: Benxi, Matang, in a temperate forest with Fagaceae, altitude 432 m, 21 August 2015, Qing Cai 1391 (HKAS 90776); same location, in a temperate forest with Fagaceae, altitude 432 m, 22 August 2015, Qing Cai 1403 (HKAS 90777). SHAANXI PROVINCE: Mei County, Taibaishan, in a temperate forest, altitude 1500 m, 4 September 2010, Xiang-Hua Wang 2741 (HKAS 61937). SHANDONG PROVINCE: Qingzhou, Yangtianshan, in a temperate forest, altitude 600 m, 22 August 2011, Xiang-Hua Wang 3012 (HKAS 77338); Taishan, in a temperate forest, altitude 1500 m, 1 August 2012, Bang Feng 1208 (HKAS 82350); Zhenping County, in a temperate forest, altitude 1535 m, 4 July 2013, Qi Zhao 1866 (HKAS 80976). YUNNAN PROVINCE: Gongshan County, Gaoligongshan, in a forest with Fagaceae, altitude 1560 m, 2 August 2010, Li-Ping Tang 1124 (HKAS 75783). JAPAN. HOKKAIDO: Tomakomai, in a forest with Quercus and Picea, altitude 100 m 12 September 2011, Zhu L. Yang 5538 (HKAS 71023).

Specimens of A. bisporigera examined: USA. TENNESSEE: Sevier County, GSMNP, ca. Gatlinburg, Cherokee Orchard [35.6811° N/83.4625° W, 945 m], 12 July 2004, mycoblitz crew s.n. [Tulloss 7-12-04-E] (HKAS 77811, RET 377-9). VIRGINIA: Giles County, near Mountain Lake, under trees of Quercus, Castanea, Betula and Acer, altitude unknown, 22 July–25 August 1946, R. Singer s.n. (FH, identified as Amanita bisporigera by R. Singer).

Commentary: Amanita pallidorosea was described from China by Zhang et al. (2010), and has also been collected from India, Japan, Pakistan, the Republic of Korea and the Russian Far East (Kim et al. 2013a; Malysheva et al. 2014; Bhatt et al. 2017; Kiran et al. 2018; this study). This species has caused at least five deaths (Zhang et al. 2010; Cao et al. 2011; Chen 2014; Chen et al. 2016), and high levels of cyclopetide toxins have been detected in the basidioma of A. pallidorosea (Li et al. 2014a; Tang et al. 2016).

In our multi-locus phylogenetic data (Fig. 2), collections labeled as A. pallidorosea split into two subclades based on GCPSR analysis. Because the differences between individuals in these two subclades were only subtle, we treat them here as a single species, A. pallidorosea. Phylogenetically, A. pallidorosea was closely related to A. bisporigera G. F. Atk (Fig. 1b). However, specimens of A. bisporigera from the USA have a non-umbonate pileus, 2-spored basidia and relatively larger basidiospores (8.0–9.5 × 7.5–9.0 μm) (Lewis 1906; Gilbert 1940, 1941a; Jenkins 1978, 1986; Tulloss et al. 1995; this study). For detailed descriptions, comparisons with similar species and images of A. pallidorosea see Zhang et al. (2010), Yang (2015) and Cai et al. (2016).

92. Amanita parviexitialis Qing Cai, Zhu L. Yang & Yang-Yang Cui, Mycologia 108 (5): 1001 (2016).

Pileus 1–3 cm in diam., slightly brownish (4B3–5) at center, becoming white (1A1) to dirty white (4A2, 4B2) towards margin; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white, unchanging. Lamellae white (11A1); lamellulae attenuate. Stipe 5–7 cm long × 0.2–0.5 cm diam., white (1A1), glabrous or sometimes covered with fibrous, white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1 cm diam.; volva shortly limbate, membranous, with free limb ca. 1 cm in height, both surfaces white (1A1). Annulus subapical, white (1A1), persistent.

Basidia (20–) 30–40 × 8–11 μm, clavate, 2-spored. Basidiospores [100/2/2] (6.5–) 7.5–9.5 (–11.0) × (6.5–) 7.0–9.0 (–11.0) μm, Q = 1.0–1.17 (–1.4), Qm = 1.09 ± 0.05, subglobose, rarely globose to broadly ellipsoid, amyloid. Interior of volval remnants on stipe base composed of irregularly, compactly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary or gregarious in subtropical forests dominated by Fagaceae; basidioma occurring in summer to autumn.

Distribution: Known from southern China (Cai et al. 2016; this study).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 480 m, 16 August 2012, Fang Li 831 [TYPE, HKAS 79049, as Amanita cf. exitialis in Yang (2015)]; same location, in a subtropical forest dominated by Fagaceae, altitude 500 m, 1 June 2013, Qing Cai 922 (HKAS 79601).

Commentary: Amanita parviexitialis corresponds to A. cf. exitialis in Yang (2015) and was later described from China by Cai et al. (2016). It is a member of the lethal Amanita subclade in A. sect. Phalloideae. Our phylogenetic study did not suggest any species that were closely related species to A. parviexitialis (Fig. 1b). For detailed descriptions, comparisons with similar species and images of A. parviexitialis see Yang (2015) as A. cf. exitialis and Cai et al. (2016).

93. Amanita pseudogemmata Hongo, Bull. Soc. Linn. Lyon (Num. spéc.): 189 (1974).

Pileus 4–9 cm in diam., dirty yellow (3A2–5) to pale yellow-brown (4B3–6); volval remnants on pileus minutely verrucose to pulverulent, brown-gray (3C3–6), brown-yellow (3B5–8) to olive-brown (2C5–8); margin striate (0.2–0.4 R), non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1) to cream (1A2); lamellulae mostly truncate. Stipe 6–10 cm long × 0.5–1.5 cm diam., cream (1A2), dirty yellow (3A2–5) to pale yellow-brown (4B3–6), covered with yellow (2A4–6) to brown-yellow (3B5–8) squamules; context white (1A1), unchanging; basal bulb sublimbate to marginate, ca. 1.5–4 cm diam., with its upper edge forming a shortly limbate, white (1A1) to yellowish (1A3–5) volval limb. Annulus superior, white (1A1) to yellowish (1A3–5) at upper surface, yellowish (1A3–5) at lower surface, persistent.

Basidia 30–50 × 10–14 μm, clavate, 4-spored. Basidiospores [150/7/7] 7.0–9.5 × 6.0–8.5 μm, Q = 1.06–1.27, Qm = 1.13 ± 0.07, subglobose to broadly ellipsoid, weakly amyloid. Volval remnants on pileus composed of more or less vertically arranged elements: filamentous hyphae fairly abundant; inflated cells abundant. Volval limb on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant; inflated cells scarce to locally abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in broad-leaved forests with Fagaceae; basidioma occurring from summer to autumn.

Distribution: Known from central, southern and southwestern China (Yang 2005, 2015; this study), and Japan (Hongo 1974b; Yang and Doi 1999).

Specimens examined: CHINA. GUANGDONG PROVINCE: Chebaling, altitude unknown, 15 July 2008, Tai-Hui Li et al. s.n. (GDGM 25449). HUBEI PROVINCE: Yichang, Shennongjia, altitude 750 m, 4 September 1984, Xiao-Qing Zhang and Shu-Xiao Sun 413 (HMAS 57649). YUNNAN PROVINCE: Longling County, in a subtropical broad-leaved forest with Lithocarpus and Castanopsis, altitude 1700 m, 12 September 2002, Zhu L. Yang 3583 (HKAS 41111); Puer, in a subtropical broad-leaved forest with Fagaceae, altitude 1326 m, 11 July 2014, Li-Hong Han 448 (HKAS 84744); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 1326 m, 11 July 2014, Jing Li 65 (HKAS 85889); Xishuangbanna, in a subtropical broad-leaved forest with Fagaceae, altitude 1130 m, 9 July 2014, Xiao-Bin Liu 413 (HKAS 87060). JAPAN. HONSHU: Kyoto, Sakyo-Ku, Ginkakuji, in a forest with Castanopsis cuspidata, 9 August 1970, H. Noro s.n. [T. Hongo 420] (Holotype, TNS-F-237280).

Commentary: Amanita pseudogemmata was described from Japan and treated as a species of A. sect. Amanita due to its striate pileal margin, pulverulent volval remnants on the pileus, a marginate to sublimbate stipe base, and the absence of clamps (Hongo 1974b; Yang 2005, 2015). However, our molecular phylogenetic analysis (Fig. 1b) indicates that it should be in A. sect. Phalloideae having amyloid basidiospores. Careful reexamination of the collections cited above showed that the basidiospores are indeed amyloid, but only weakly.

In our phylogenetic analysis inferred from ITS data (Fig. S11), A. pseudogemmata is sister to A. levistriata. However, the latter differs morphologically in its relatively smaller basidioma with a ca. 1.7–3 cm in diam. pileus, volval remnants on the pileus as floccose patches at center, becoming flocculose towards the margin, and its occurrence in sandy soils in mixed coniferous and deciduous forests (Jenkins 1988). For detailed descriptions, comparisons with similar species and images of pseudogemmata see Yang (2005, 2015).

94. Amanita rimosa P. Zhang & Zhu L. Yang, Fungal Divers. 42: 124 (2010).

Pileus 3–5 cm in diam., cream (1A2) to pale yellow-brown (1B2–4) at center, becoming white (1A1) at margin, often slightly rimose at margin; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 5–8 cm long × 0.3–1 cm diam., white (1A1) to dirty white (2B1), sometimes covered with minute, white (1A1) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 0.8–1.6 cm diam.; volva limbate, membranous, with free limb ca. 0.8 cm in height, both surfaces white (1A1). Annulus subapical, white (1A1), persistent. All parts of basidioma becoming yellow (2A5–8) immediately with 5% KOH solution.

Basidia (20–) 35–48 × 10–13 μm, clavate, 4-spored. Basidiospores [260/15/13] 7.0–8.5 (–10.0) × 6.5–8.0 (–9.0) μm, Q = (1.0–) 1.05–1.15 (–1.23), Qm = 1.08 ± 0.05, globose to subglobose, amyloid. Pileipellis composed of two layers: upper layer slightly gelatinized, composed of radially arranged, ellipsoid to clavate inflated cells, mixed with filamentous hyphae; lower layer composed of radially and compactly arranged filamentous hyphae. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in tropical to subtropical broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring from late spring to autumn.

Distribution: Known in eastern, central and southern China (Zhang et al. 2010; Yang 2015; Cai et al. 2016; this study), and Thailand (Thongbai et al. 2016).

Specimens examined: CHINA. GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 450 m, 17 May 2012, Fang Li 302 (HKAS 101393); Guangzhou, Baiyunshan, in a subtropical broad-leaved forest with Fagaceae, altitude 190 m, 28 May 2013, Qing Cai 891 (HKAS 79570); same location, in a subtropical broad-leaved forest with Fagaceae, altitude 380 m, 28 May 2013, Yan-Jia Hao 804 (HKAS 80084); Guangzhou, Tianluhu, in a subtropical broad-leaved forest with Castanopsis and Lithocarpus, altitude 400 m, 29 May 2013, Qing Cai 904 (HKAS 79583); Shixing County, in a forest of Castanopsis, altitude 365 m, 18 September 2011, Xiang-Hua Wang 3179 (HKAS 75779); Zhaoqing, Dinghushan, in a subtropical forest dominated by Fagaceae, altitude 200 m, 9 August 2010, Nian-Kai Zeng 874 (HKAS 77336). HAINAN PROVINCE: Baisha County, Yinggeling, in a tropical to subtropical forest, altitude 1790 m, 26 July 2009, Nian-Kai Zeng 329 (HKAS 75777); Qiongzhong County, Limushan, in a tropical to subtropical forest, altitude 170 m, 23 July 2009, Nian-Kai Zeng 301 (HKAS 75778); same location, in a tropical to subtropical forest, altitude 170 m, 20 May 2009, Nian-Kai Zeng 186 (HKAS 77335). HUNAN ROVINCE: Yizhang County, Mangshan, in a subtropical mixed forest with Fagaceae and Pinaceae, altitude 1300 m, 10 September 2005, Ping Zhang 487 (TYPE, HKAS 49675). JIANGXI PROVINCE: Fuzhou, in a subtropical forest dominated by Fagaceae, altitude 60 m, 20 June 2012, Gang Wu 948 (HKAS 77120); same location, in a forest dominated by Fagaceae, altitude 60 m, 19 June 2012, Gang Wu 933 (HKAS 77105); Linchuan, Zhanping, in a subtropical forest, altitude 140 m, 24 June 2009, Hui Cheng 090624 (HKAS 58667).

Commentary: Amanita rimosa was described from China by Zhang et al. (2010), and subsequently reported from Thailand (Thongbai et al. 2016). This species belongs to the lethal Amanita subclade of A. sect. Phalloideae, it contains the highest toxin content of amatoxins and phallotoxins of the Chinese lethal Amanita (Tang et al. 2016). During 2014–2016, five known cases of mushroom poisoning caused by A. rimosa occurred in eastern and southern China (Chen et al. 2016; Jiang et al. 2016).

Our multi-locus phylogenetic analysis (Fig. 1b) did not suggest any species related to Amanita rimosa within A. sect. Phalloideae. For detailed descriptions, comparisons with similar species and images of A. rimosa see Zhang et al. (2010), Yang (2015) and Cai et al. (2016).

95. Amanita subfuliginea Qing Cai, Zhu L. Yang & Yang-Yang Cui, Mycologia 108 (5): 1003 (2016).

Pileus 4–5.5 cm in diam., brown (4D3–6) to dark brown (4E2–6), with innate, brown to dark brown, radiating fibrils; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 10–12 cm long × 0.5–0.7 cm diam., grayish (4C2–4) to brownish (4B2–4), densely covered with brown (4D4–6) to dark brown (4E5–8) fibrillose squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1–1.3 cm diam.; volva limbate, membranous, with free limb ca. 1 cm in height, both surfaces white (1A1). Annulus apical to subapical, white (1A1), persistent.

Basidia 30–45 × 8–11 μm, clavate, 4-spored. Basidiospores [90/5/3] (6.0–) 7.0–9.0 (–11.0) × (6.0–) 7.0–9.0 (–10.5) μm, Q = 1.0–1.03 (–1.06), Qm = 1.01 ± 0.02, globose, rarely subglobose, amyloid. Interior of volval remnants on stipe base composed of longitudinally arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical pine or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer to autumn.

Distribution: Known in central and southern China (Cai et al. 2016; this study).

Specimens examined: CHINA. GUANGDONG PROVINCE: Lechang, Yangdongshan, in a subtropical forest dominated by Pinus kwangdongensis, altitude 1200 m, 15 September 2011, Xiang-Hua Wang 3148 (TYPE, HKAS 77326). HUNAN PROVINCE: Yizhang County, Mangshan, in a subtropical forest, altitude 1300 m, 9 September 2005, Ping Zhang 460 (HKAS 77347); same location, in a subtropical mixed forest dominated by Pinus massoniana, altitude 1300 m, 29 September 1981, Yu-Chen Song and Xiao-Lan Mao 63 (HMAS 42198).

Commentary: Amanita subfuliginea corresponds to Amanita sp. 4 in Cai et al. (2014) and was later described from China by Cai et al. (2016). This species is a member of the lethal Amanita subclade of A. sect. Phalloideae. Phylogenetically, A. subfuliginea is closely related A. fuliginea (Fig. 1b), but A. fuliginea has a paler stipe (white, gray to brownish), and globose to subglobose basidiospores (Hongo 1953, Yang 1997, 2005, 2015, Cai et al. 2016; Imazeki et al. 2011; this study). For detailed descriptions, comparisons with similar species and images of A. subfuliginea see Cai et al. (2016).

96. Amanita subjunquillea S. Imai, Bot Mag (Tokyo) 47: 424 (1933).

Synonym: Amanita subjunquillea var. alba Zhu L. Yang, Bibl. Mycol. 170: 174 (1997), syn. nov.

Pileus 3–6 cm in diam., sometimes umbonate at center, yellow-brown (1B4–7), dirty orange-yellow (5A2–5) to mustard yellow (30A2–4), sometimes entire white (1A1); volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 4–12 (–15) cm long × 0.3–1 (–1.5) cm diam., white (1A1) to yellowish (1A2–4), covered with fibrous, yellowish (1A2–4) squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1–2 (–2.5) cm diam.; volva limbate, membranous, with free limb up to 2 cm in height, both surfaces white (1A1) to dirty white (2B1). Annulus subapical to superior, white (1A1), persistent or fragile and fugacious. All parts of entirely white basidioma becoming yellow (2A5–8) immediately with 5% KOH solution.

Basidia 30–48 × 9–12 μm, clavate, 4-spored. Basidiospores [290/42/39] 6.5–9.5 (–11.0) × (5.5–) 6.0–8.0 (–10.0) μm, Q = (1.0–) 1.03–1.19 (–1.23), Qm = 1.1 ± 0.05, globose to subglobose, sometimes broadly ellipsoid, amyloid. Interior of volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical to temperate forests dominated by Fagaceae; basidioma occurring from summer to autumn.

Distribution: Known from central, northern, northeastern, northwestern and southwestern China (Yang 1997, 2005, 2015; Cai et al. 2016; this study), India (Bhatt et al. 2003, 2007), Japan (Imai 1933, 1938; Yang and Doi 1999; Imazeki et al. 2011) and Republic of Korea (Kim et al. 2013a; Cho et al. 2015).

Specimens examined: CHINA. ANHUI PROVINCE: Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 1195 m, 20 July 2017, Yan-Jia Hao 1480 (HKAS 100597); same location, in a forest with Fagaceae, altitude 1100 m, 19 July 2017, Yan-Jia Hao 1439 (HKAS 100581); same location, in a forest with Fagaceae and Pinaceae, altitude 955 m, 23 July 2017, Yan-Jia Hao 1545 (HKAS 100622). GANSU PROVINCE: Tianshui, Maijishan, altitude 1800 m, 4 August 2012, Qi Zhao 1627 (HKAS 75772); Tianshui, Taohuagou, altitude 1300 m, 25 July 2011, Xiao-Fei Shi 745 (HKAS 101428). GUIZHOU PROVINCE: Daozhen County, in a subtropical forest, altitude 1350 m, 19 July 1988, Jian-Zhe Ying, Yu-Chen Song and Yu Li G405 (HMAS 59783); same location, in a subtropical forest, altitude 1350 m, 20 July 1988, Jian-Zhe Ying, Yu-Chen Song and Yu Li 475 (HMAS 60316); same location, in a subtropical forest, altitude 1350 m, 20 July 1988, Jian-Zhe Ying, Yu-Chen Song and Yu Li 401 (HMAS 59785). HENAN PROVINCE: Neixiang, Taohuacun, altitude 180 m, 31 July 2010, Xiang-Hua Wang 2441 (HKAS 77325). HUBEI PROVINCE: Fang County, in a forest with Fagaceae, altitude 1200 m, 31 July 2011, Xiao-Fei Shi 768 (HKAS 77345); Yichang, Shennongjia, in a forest with Fagaceae, altitude 1840 m, 10 August 2015, Yang-Yang Cui 263 (HKAS 89695); same location, in a forest with Fagaceae, altitude 1780 m, 11 August 2015, Yang-Yang Cui 267 (HKAS 89696); same location, in a forest with Fagaceae, altitude 1770 m, 11 August 2015, Yang-Yang Cui 268 (HKAS 89697); same location, in a forest with Fagaceae, altitude 1780 m, 12 August 2015, Yang-Yang Cui 270 (HKAS 89698); same location, in a forest with Fagaceae, altitude 1770 m, 12 August 2015, Yang-Yang Cui 271 (HKAS 89699); same location, in a forest with Quercus, altitude 1850 m, 15 July 2012, Qing Cai 796 (HKAS 75550); same location, in a forest with Quercus, altitude 1850 m, 15 July 2012, Qing Cai 798 (HKAS 75552); same location, in a forest with Fagaceae, altitude 1900 m, 18 July 2012, Qing Cai 845 (HKAS 75771). INNER MONGOLIA AUTONOMOUS REGION: Daqinggou, in a temperate forest, altitude 270 m, 24 August 2005, Tolgor Bau 5788 (HMJAU 57882). JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest, altitude 680 m, 7 August 2010, Xiao-Fei Shi 453 (HKAS 63418); Fusong County, Lushuihe, in a temperate forest, altitude 740 m, 3 October 2001, Tolgor Bau 6772 (HMJAU 6772). SHAANXI PROVINCE: Mei County, Haoping, altitude 1235, 5 September 2010, Xiang-Hua Wang 2753 (HKAS 75770). LIAONING PROVINCE: Benxi, Matang, in a temperate forest with Fagaceae, altitude 432 m, 21 August 2015, Qing Cai 1385 (HKAS 90778); Anshan City, Qianshan, in a temperate mixed forest with Fagaceae and Pinus tabuliformis, altitude 140 m, 24 August 2015, Qing Cai 1414 (HKAS 90779). SHAANXI PROVINCE: Zhouzhi County, Taibaishan, in a forest with Abies, altitude 1600 m, 27 July 2011, Xiao-Fei Shi 755 (HKAS 101429). YUNNAN ROVINCE: Lanping County, in a subtropical mixed forest with Fagaceae and Pinus armandii, altitude 2700 m, 14 August 2011, Gang Wu 679 (HKAS 74993); Kunming, Yeyanhu, in a forest with Quercus variabilis, altitude 1990 m, 17 September 2008, Zhu L. Yang 5213 (HKAS 54509). JAPAN. HONSHU: Tokyo, Oume, Kurosawa 3-Chôme, left-side ridge of Kurosawa-gawa River, along hiking course, 16 July 1997, Y. Doi & K. Miyazaki s.n. (TNS-F-182483); Tokyo, Oume, Kurosawa 3-Chôme, right-side ridge of Kurosawa-gawa R., 25 September 1997, Y. Doi s.n. (TNS-F-237829); Tokyo, Oume, Kurosawa 3-Chôme, Nagayama-Kyûryô Hills, along hiking course, 5 October 1997, Y. Doi s.n. (TNS-F-237728).

Specimens (albino form) examined: CHINA. GUIZHOU PROVINCE: Daozhen County, Dashahe, in a mixed forest, altitude unknown, 20 July 1988, Jian-Zhe Ying et al. 471 (HMAS 58289). SICHUAN PROVINCE: Xichang County, Luojishan, altitude 2100 m, 18 July 1983, Ming-Sheng Yuan 31 (HKAS 11831). TIBET AUTONOMOUS PREFECTURE: Chayu County, altitude ca. 2300 m, 1 September 1976, Mu Zang 664 (HKAS 5664). YUNNAN PROVINCE: Menghai County, in a subtropical forest with Pinus kesiya var. langbianensis, altitude 1200 m, 14 August 1991, Zhu L. Yang 1530 (HKAS 24169). JAPAN. HONSHU: Chiba, Kiyosumi, Fudagou, Do-zawa, 14. July 1983, Y. Doi s.n. (TNS-F-174870); Kyoto, Ahiu, Kyoto Univ. For., 24 July 1964, Y. Doi s.n. (TNS-F-194890); Tokyo, Oume, Kurosawa 3-Chôme, left-side ridge of Kurosawa-gawa River, along hiking course, 5 October 1997, Y. Doi s.n. (TNS-F-237599); Tokyo, Oume, Kurosawa 3-Chôme, left-side ridge of Kurosawa-gawa River, along hiking course, 5 October 1997, Y. Doi s.n. (TNS-F-237594); Tokyo, Oume City, Kurosawa 3-Chôme, right-side ridge of Kurosawa-gawa River, 25 September 1997, Y. Doi s.n. (TNS-F-237846).

Specimens of A. phalloides examined: GERMANY. BADEN-WURTTEMBERG, Tübingen, under Fagus sylvatica, altitude ca. 400 m, 3 September 1994, Zhu L. Yang 30D (HKAS 31625); same location, 10 September 1994, Zhu L. Yang 32D (HKAS 31457).

Commentary: Amanita subjunquillea was described from Japan by Imai (1933), and then reported from China, India and Republic of Korea (Bhatt et al. 2003, 2007; Yang 1997, 2005, 2015; Kim et al. 2013a; Cho et al. 2015; Cai et al. 2016). This species is deadly poisonous (Kawase et al. 1992; Imazeki et al. 1988; Chen et al. 2016; Tang et al. 2016). The albino form of A. subjunquillea was treated as A. subjunquillea var. alba by Yang (1997, 2005, 2015). Since few genetic differences between the albino form and the non-albino form are present, we treat A. subjunquillea var. alba as a synonym of A. subjunquillea var. subjunquillea (Cai et al. 2014).

Phylogenetically, A. subjunquillea is closely related to A. phalloides (Vaill.:Fr.) Link (Fig. 1b). Indeed, they are also morphologically similar to each other. However, A. phalloides differs from A. subjunquillea in its robust basidioma and subglobose to broadly ellipsoid basidiospores (8.0–10.5 × 6.5–9.0 μm, Q = 1.1–1.29, Qm = 1.19 ± 0.06) (Gilbert 1940, 1941a; Thiers 1982; Contu 2000a; Neville and Poumarat 2004; this study). For detailed descriptions, comparisons with similar species and images of A. subjunquillea see Yang (1997, 2005, 2015) and Cai et al. (2016).

97. Amanita subpallidorosea Hai J. Li, Mycol. Prog. 14 (6/43): 5 (2015).

Pileus 5–8 cm in diam., umbonate at center, white (1A1), often pinkish (7A2–4) at center; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 7–10 cm long × 0.6–1.2 cm diam., white (1A1) to dirty white (2B1), covered with concolorous squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 1.5–2.5 cm diam.; volva limbate, membranous, with free limb up to 1 cm in height, both surfaces white (1A1). Annulus subapical, white (1A1), persistent. All parts of entirely white basidioma becoming yellow (2A5–8) immediately with 5% KOH solution.

Basidia 35–50 × 11–13 μm, clavate, 4-spored. Basidiospores [20/1/1] (7.5–) 8.0–10.0 (–11.5) × (7.0–) 7.5–9.0 (–10.0) μm, Q = (1.0–) 1.02–1.11 (–1.13), Qm = 1.07 ± 0.03, globose to subglobose, amyloid. Interior of volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical forests dominated by Quercus and Cyclobalanopsis; basidioma occurring in summer to autumn.

Distribution: Known from eastern and southwestern China (Li et al. 2015; Yang 2015; Cai et al. 2016; this study).

Specimen examined: CHINA. TAIWAN ROVINCE: Taizhong, altitude ca. 2000 m, 19 September 2012, Bang Feng 1324 (HKAS 77350).

Commentary: Amanita subpallidorosea was described from China by Li et al. (2015) and is a member of the lethal Amanita subclade of A. sect. Phalloideae. This lethal Amanita species is known to have caused two deaths in Guizhou Province, China in 2014 (Li et al. 2015; Chen et al. 2016). Thirteen cyclopeptides were subsequently identified from the species (Wei et al. 2017).

According to our multi-locus phylogenetic analysis (Fig. 1b), collection HKAS 27133, which was identified as A. cf. virosa by Weiß et al. (1998), clustered within A. subpallidorosea and should be considered A. subpallidorosea. Phylogenetically, A. subpallidorosea is related to A. virosa Bertillon (Fig. 1b). However, A. virosa has a stipe covered with recurved squamules, and larger basidiospores (Contu 2000a; Chiusa 2000; Neville and Poumarat 2004; Zhang et al. 2010; Li et al. 2015; Yang 2015; Cai et al. 2016; this study). Furthermore, A. virosa is dominant in temperate Eurasia (Contu 2000a; Chiusa 2000; Neville and Poumarat 2004; Zhang et al. 2010; Li et al. 2015; Yang 2015; Cai et al. 2016; this study). For detailed descriptions, comparisons with similar species and images of A. subpallidorosea see Li et al. (2015), Yang (2015) and Cai et al. (2016).

98. Amanita virosa Bertillon, Dict. Encycl. Sci. Médic.: 497 (1866).

Replaced synonym: Agaricus virosus Fr., Epicr. syst. mycol. (Upsaliae): 3 (1838) [1836-1838]; non Agaricus virosus Sowerby, Col. fig. Engl. Fung. Mushr. (London) 3: tab. 407 (1809).

Pileus 5–10 cm in diam., umbonate at center, white (1A1), often cream (1A2) at center; volval remnants on pileus absent; margin non-striate, non-appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 8–16 cm long × 0.7–2 cm diam., white (1A1), covered with concolorous squamules; context white (1A1), unchanging; basal bulb subglobose, ca. 2–3 cm diam.; volva limbate, membranous, with free limb up to 1 cm in height, both surfaces white (1A1). Annulus subapical, white (1A1), persistent. All parts of basidioma becoming yellow (2A5–8) immediately with 5% KOH solution.

Basidia 30–50 × 10–12 μm, clavate, 4-spored. Basidiospores [100/9/9] (7.5–) 8.0–11.0 (–12.0) × (7.0–) 8.0–10.0 (–11.0) μm, Q = 1.0–1.14 (–1.21), Qm = 1.07 ± 0.05, globose to subglobose, amyloid. Interior of volval remnants on stipe base composed of longitudinally to irregularly arranged elements: filamentous hyphae very abundant to nearly dominant; inflated cells scarce to scattered. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in subtropical to temperate forests associated with Fagaceae and Pinaceae; basidioma occurring from summer to autumn.

Distribution: Known in Europe (Konrad and Maublanc 1924; Contu 2000a; Chiusa 2000; Neville and Poumarat 2004) and East Asia (Imai 1933, 1938; Zhang et al. 2010; Imazeki et al. 2011; Li et al. 2015, Yang 2015; Cai et al. 2016).

Specimens examined: CHINA. JILIN PROVINCE: Antu County, Changbaishan, in a temperate forest, altitude ca. 1000 m, 9 September 2006, Xiang-Hua Wang 2071 (HKAS 50912); same location, in a temperate forest, altitude 1162 m, 30 August 2015, Qing Cai 1470 (HKAS 90176); same location, in a forest dominated by Fagaceae, altitude 1400 m, 14 September 2014, Xiao-Bin Liu 712 (HKAS 87260). HUNAN PROVINCE: Badagongshan, in a forest with Fagaceae, 9 September 2016, Ping Zhng 2121 (MHHNU 8621). AUSTRIA. VORARLBERG: Kleinwalsertal, altitude 1150 m, 20 September 2009, KR 9741 (MB-001176). FINLAND. EZASTERN FINLAND PROVINCE: Juva, under Pinus, mixed with Picea and Abies, altitude ca. 120 m, date unknown, Xiang-Hua Wang 2293 (HKAS 56694). JAPAN. HOKKAIDO: Tomakomai, in a forest under Quercus, altitude 100 m, 13 September 2011, Zhu L. Yang 5555 (HKAS 71040). POLAND. WOJEWÓDZTWO PODLASKIE: Białowieża Forest, altitude unknown, 5 September 1966, C. Bas 4697 (L). SWEDEN. DALARNA: Bjursås, in a pine forest, altitude unknown, 28 August 1982, R. Morander 4343 (F 4656).

Commentary: Amanita virosa is a member of the lethal Amanita subclade in A. sect. Phalloideae. This species has a wide distribution across Europe and temperate to subtropical Asia (Contu 2000a; Chiusa 2000; Neville and Poumarat 2004, Zhang et al. 2010, Li et al. 2015, Yang 2015; Cai et al. 2016; this study). Cyclopetide toxins have been detected in basidiomata collected from Europe (Wieland et al. 1978; Faulstich et al. 1980; Buku et al. 1980a, b; Wienland and Faulstich 1991; Antonyuk et al. 2010; Wieland 2012) and from East Asia (Li et al. 2014a; Tang et al. 2016; Wei et al. 2017).

Our multi-locus phylogenetic analysis (Fig. 1b) indicated that A. virosa is closely related to A. subpallidorosea, and the comparisons between them were presented in the commentary on A. subpallidorosea. For detailed descriptions, comparisons with similar species and images of A. virosa see Yang (2015).

99. Amanita zangii Zhu L. Yang, T. H. Li & X. L. Wu, Fungal Divers. 6: 160 (2001).

Synonym: Amanita areolata T. Oda, C. Tanaka & Tsuda, Mycoscience 43(4): 351 (2002), syn. nov.

Pileus 5–6 cm in diam., white (1A1) to cream (1A2); volval remnants on pileus subconical to flat verrucose, felty to subfibrillose, brown-gray (2E4–6), dark gray (1E1–3) to blackish (4F1–3); margin non-striate, appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 6–8 cm long × 0.8–1.2 cm diam., white (1A1) to cream (1A2), covered with fibrous to floccose, concolorous squamules; context white (1A1), unchanging; basal bulb subclavate, occasionally subglobose, ca. 1.5–2 cm diam., with its upper part covered with pulverulent to floccose, white (1A1) to gray (3B1) volval remnants. Annulus apical to subapical, white (1A1), fragile and fugacious.

Basidia 30–50 × 9.5–12 μm, clavate, 4-spored. Basidiospores [60/3/3] (7.5–) 8.5–11.5 (–12.0) × 6.5–8.0 (–9.0) μm, Q = (1.25–) 1.29–1.57 (–1.69), Qm = 1.41 ± 0.09, ellipsoid, amyloid. Volval remnants on pileus composed of more or less vertically arranged elements: filamentous hyphae fairly abundant to scattered; inflated cells very abundant. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered on soil in tropical to subtropical forests; basidioma occurring from summer to autumn.

Distribution: Known in southern China (Yang et al. 2001; Yang 2005, 2015; this study), Japan (Oda et al. 2002a, identified as A. areolata) and Thailand (Thongbai et al. 2016, 2017).

Specimens examined: CHINA. HAINAN PROVINCE: Qiongzhong County, Limushan, in a tropical to subtropical forest, altitude 750 m, 4 August 2010, Nian-Kai Zeng 835 (HKAS 77331); same location, in a forest dominated by Fagaceae, altitude 840 m, 21 June 2016, Gang Wu 1546 (HKAS 99663); Ledong County, Jianfengling, in a tropical to subtropical forest, altitude 900 m, 20 August 1999, Ming-Sheng Yuan 4346 (HOLOTYPE, HKAS 34570).

Commentary: Amanita zangii was placed in A. sect. Lepidella sensu Bas (1969) exclusively based on morphological study, due to the felted to subfibrillose volval remnants on the pileus, appendiculate pileal margins, narrowly clavate to subfusiform stipe bases, and the volval remnants on the pileus with long ellipsoid to subfusiform inflated cells (Yang et al. 2001; Yang 2005, 2015). It was suggested that this species is very similar to A. hesleri and A. inopinata D. A. Reid & Bas (Yang et al. 2001). Our molecular phylogenetic analysis (Fig. 1b) indicates that it is indeed sister to A. hesleri Bas, but that it should be moved to A. sect. Phalloideae (Cai et al. 2014; this study).

Amanita areolata T. Oda et al., described from Japan, was suspected to be a synonym of A. zangii (Yang 2005). Phylogenetically, the type of A. areolata fell within the clade of A. zangii with high support (Cai et al. 2014), and the morphological characters of A. areolata are identical to those in A. zangii. Therefore, we treat A. areolata as a synonym of A. zangii. For detailed descriptions, comparisons with similar species and images of A. zangii see Yang et al. (2001) and Yang (2005, 2015).

Amanita sect. Roanokenses Singer ex Singer, Sydowia 15: 67 (1962).

Synonym: Amanita subsect. Limbatulae Bas, Persoonia 5: 528 (1969).

Type: Amanita roanokensis Coker, J. Elisha Mitchell scient. Soc. 43: 141 (1927).

Diagnosis: Pileus white, dirty white, grayish, gray to brownish; pileal margin non-striate, appendiculate; lamellulae attenuate; stipe with bulbous base; volval remnants often as cones, verrucae, pyramids, powder, floccules to patches on pileal surface and stipe base, occasionally absent on pileus and then leaving a limbate volva on stipe base; annulus membranous to fugacious; basidiospores amyloid; clamps present or absent.

Commentary: Our multi-locus phylogenetic analysis (Fig. 1a, b) indicates that Amanita sect. Lepidella sensu Bas (1969) is polyphyletic and clusters in four clades across two subgenera: sect. Lepidella of Amanita subgen. Lepidella; sect. Strobiliformes, sect. Phalloideae and sect. Roanokenses of Amanita subgen. Amanitina. Amanita sect. Roanokenses was proposed by Singer in 1951 but was invalid due to the absence of the Latin diagnosis (Bas 1969; Neville and Poumarat 2004). Singer (1962a) then effectively proposed the name of A. sect. Roanokenses Singer, typified by A. roanokensis Coker, and provided a brief Latin diagnosis, “Pileo stipiteque pulvere detersili obtectis”. In this study, 35 species of A. sect. Roanokenses from China are recorded below, including 13 novel species and 22 known ones.

Key to the species of Amanita sect. Roanokenses from China

  1. 1.

    Volval remnants often as pyramids, warts, flocci, powder or patches persistent on pileus and stipe base……….2

     
  2. 1.

    Volval remnants membranous, and often attached to bulb as limb, occasionally with flat patches retained on pileus……….29

     
  3. 2.

    Lamellae white, cream or yellow; odor indistinct when dried……….3

     
  4. 2.

    Lamellae biscuit to flesh colored; odor like tomato juice when dried………A. avellaneifolia

     
  5. 3.

    Basidioma with color change when injured……….4

     
  6. 3.

    Basidioma lacking color change when injured……….8

     
  7. 4.

    Basidioma becoming pinkish, brownish, brown, dark brown to black when injured………

     
  8. 4.

    Basidioma becoming yellow when injured……….7

     
  9. 5.

    Basidioma becoming pinkish to brown when injured; basidiospores broadly ellipsoid to ellipsoid, 5.5–6.5 μm in width…………….6

     
  10. 5.

    Basidioma becoming brownish, dark brown to black when injured; basidiospores globose to subglobose, 8.0–10.5 μm in width………A. sculpta

     
  11. 6.

    Volval remnants on stipe base often absent; basidiospores subglobose to broadly ellipsoid, 6.0–8.5 × 5.5–7.5 μm………A. gymnopus

     
  12. 6.

    Volval remnants on stipe base present and conical; basidiospores ellipsoid, 8.5–10.0 × 5.5–6.5 μm………A. pannosa

     
  13. 7.

    Basidioma small to medium-sized with pileus 4–9 cm in diam.; pileus white to dirty white covered with concolorous, floccose, conical to patchy volval remnants; stipe white covered with concolorous, minute squamules; volval remnants on stipe base patchy; basidiospores ellipsoid to elongate, 8.0–11.0 × 4.5–5.5 μm, Q = 1.45–2.0; clamps absent; odor unpleasant………A. alboflavescens

     
  14. 7.

    Basidioma very large with pileus 15–24 cm in diam.; pileus brownish with orange tinge, and covered with dirty yellow to orange, pyramidal volval remnants; stipe yellowish covered with yellowish to brownish, snakeskin-shaped squamules; volval remnants on stipe base verrucose; basidiospores ellipsoid, 7.0–9.0 × 5.0–6.0 μm, Q = 1.3–1.7; clamps present; odor indistinct………A. macrocarpa

     
  15. 8.

    Both pileus and volval remnants on its surface white to dirty white, sometimes with pink or brown tinge……….9

     
  16. 8.

    Either pileus or volval remnants on its surface grayish, gray, dark gray, brownish, cameo brown or yellowish……….19

     
  17. 9.

    Pileus and volval remnants on its surface often with pink to flesh red tinge……….10

     
  18. 9.

    Pileus and volval remnants on its surface white, or sometimes with yellow tinge……….12

     
  19. 10.

    Stipe base fusiform to ovoid, not radicating; clamps present; basidiospores broadly ellipsoid to ellipsoid or ovoid……….11

     
  20. 10.

    Stipe base fusiform, strobiliform to long clavate, radicating; clamps absent; basidiospores elongate to ellipsoid………A. brunneostrobilipes

     
  21. 11.

    Basidiospores broadly ellipsoid to ellipsoid, 9.0–11.0 × 7.0–8.0 μm, Q = 1.21–1.47; basidioma distributed in subtropical to temperate pine, broad-leaved to mixed forests………A. eijii

     
  22. 11.

    Basidiospores broadly ellipsoid to ovoid, 7.0–9.5 × 5.5–7.0 μm, Q = 1.15–1.36; basidioma distributed in tropical to subtropical broad-leaved forests………A. timida

     
  23. 12.

    Clamps absent……….13

     
  24. 12.

    Clamps present……….15

     
  25. 13.

    Volval remnants on pileus white, pyramidal, not separated into two layers, dominantly composed of rare filamentous hyphae and very abundant inflated cells; volval remnants on stipe base pyramidal……….14

     
  26. 13.

    Volval remnants on pileus dirty white to cream and separated into two layers: outer layer membranous, dominantly composed of very abundant filamentous hyphae and scarce inflated cells, inner layer pulverulent, dominantly composed of scarce filamentous hyphae and very abundant inflated cells; volval remnants on stipe base patchy or arranged in incomplete belts………A. neoovoidea

     
  27. 14.

    Basidioma medium-sized to large with pileus 5–10 cm in diam.; basidiospores ellipsoid to elongate, 7.5–10.0 × 4.5–6.5 μm, Q = 1.33–1.80………A. yenii

     
  28. 14.

    Basidioma small with pileus 3–5 cm in diam.; basidiospores mostly subglobose to broadly ellipsoid, 6.0–8.5 × 6.0–7.5 μm, Q = 1.03–1.17………A. parvivirginea

     
  29. 15.

    Stipe base fusiform to napiform, occasionally subglobose; basidiospores broadly ellipsoid, ellipsoid to elongate, Q = 1.18–2.02……….16

     
  30. 15.

    Stipe base subglobose to globose; basidiospores mostly subglobose, Q  = 1.04–1.14………A. sphaerobulbosa

     
  31. 16.

    Basidioma medium-sized to large with pileus 5–15 cm in diam.; volval remnants on pileus conical to verrucose; lamellae white, cream to brownish; odor indistinct……….17

     
  32. 16.

    Basidioma small to medium-sized with pileus 4–7 cm in diam.; volval remnants on pileus felted to patchy; lamellae yellowish; odor unpleasant……………………………………A. kotohiraensis

     
  33. 17.

    Basidioma medium-sized to large; pyramids on pileus relatively smaller (≤ 3 mm in height and width); basidiospores broadly ellipsoid to ellipsoid, 7.0–10.0 × 5.0–7.5 μm, Q = 1.18–1.6……….18

     
  34. 17.

    Basidioma mostly medium-sized; pyramids on pileus relatively larger (2–5 mm in height and 2–8 mm in width); basidiospores ellipsoid to elongate, 9.0–11.0 × 5.0–6.5 μm, Q = 1.54–2.02, Qm = 1.81 ± 0.18………A. castanopsis

     
  35. 18.

    Lamellae white; basidiospores relatively broader, broadly ellipsoid to ellipsoid, 8.0–10.0 × 6.0–7.5 μm, Q = 1.18–1.50; basidioma distributed in pine, broad-leaved or mixed forests………A. virgineoides

     
  36. 18.

    Lamellae cream to brownish; basidiospores relatively narrower, mostly ellipsoid, 7.0–9.0 × 5.0–6.5 μm, Q = 1.3–1.6; basidioma distributed in tropical to subtropical broad-leaved forests………A. minutisquama

     
  37. 19.

    Clamps absent……….20

     
  38. 19.

    Clamps present……….22

     
  39. 20.

    Basidioma gray; volval remnants on pileus pulverulent, floccose to felted; stipe surface covered with tomentose, floccose to pulverulent squamules……….21

     
  40. 20.

    Basidioma dirty yellow, yellow, brownish to yellowish; volval remnants on pileus pyramidal over entire disk; stipe surface covered with conical concentrically arranged squamules………A. hongoi

     
  41. 21.

    Volval remnants on pileus tomentose to felted, yellow-brown, brownish to dark brown; basidiospores relatively narrower, 7.5–9.5 × 5.5–6.5 μm, Q = 1.33–1.58, Qm = 1.45 ± 0.09………A. vestita

     
  42. 21.

    Volval remnants on pileus tomentose, floccose to pulverulent, gray, grayish brown to dark gray; basidiospores relatively broader, 8.0–10.0 × 6.0–9.5 μm, Q = 1–1.35, Qm = 1.18 ± 0.15………A. griseofarinosa

     
  43. 22.

    Basidiospores mostly globose, subglobose to broadly ellipsoid, occasionally ellipsoid, Q = 1.0–1.43……….23

     
  44. 22.

    Basidiospores ellipsoid to elongate, Q = 1.3–1.8……….24

     
  45. 23.

    Verrucose to pyramidal volval remnants on pileus gray to dark gray, lacking white tinge; stipe base lacking red tinge; volval remnants on stipe base forming as gray to gray-brown, recurved squamules, dominantly composed of scarce filamentous hyphae and very abundant, ellipsoid to clavate inflated cells; basidioma distributed in subalpine forests with Abies and Picea………A. pyramidata

     
  46. 23.

    Verrucose to pyramidal volval remnants on pileus gray, often with apical part white; stipe base often with red tinge; volval remnants on stipe base forming as verrucae, dominantly composed of abundant filamentous hyphae and abundant, subglobose, globose, fusiform to ellipsoid inflated cells; basidioma distributed in pine, broad-leaved or mixed forests associated with Castanopsis, Lithocarpus, Quercus and Pinus………A. rubiginosa

     
  47. 24.

    Basidiospores relatively smaller, 7.0–9.0 × 5.0–6.0 μm; lamellae white, cream or greenish……….25

     
  48. 24.

    Basidiospores relatively larger, 8.5–12.0 × 5.5–7.5 μm; lamellae white, cream or yellowish to lemon yellow……….26

     
  49. 25.

    Lamellae white to cream; stipe base ellipsoid, elongate to clavate; basidioma distributed in tropical to subtropical broad-leaved forests………A. elliptica

     
  50. 25.

    Lamellae greenish; stipe base subglobose; basidioma distributed in mixed forests………A. pallidochlorotica

     
  51. 26.

    Lamellae white to cream; volval remnants on pileus pyramidal to floccose; gray, brown-gray to brown, composed of abundant globose to subglobose inflated cells; volval remnants on stipe base floccose, conical to verrucose, dirty white to gray……….27

     
  52. 26.

    Lamellae yellowish to lemon-yellow; volval remnants on pileus floccose to felted, cameo brown, composed of abundant fusiform to ellipsoid inflated cells; volval remnants on stipe base floccose to pulverulent, cameo brown to brown………A. luteofolia

     
  53. 27.

    Basidioma relatively slender with pileus ca 4–9 cm in diam.; pyramids on pileus relatively smaller, ca. 1–2 mm in height and width; basidioma fruited in summer to autumn……….28

     
  54. 27.

    Basidioma relatively robuster with pileus ca. 5.5–13.5 cm in diam.; pyramids on pileus relatively larger, ca. 1–6 mm in height and width; basidioma fruited in early spring………A. heishidingensis

     
  55. 28.

    Volval remnants on the pileus floccose to pyramidal; stipe base napiform, sometimes slightly radicating, lacking distinct pesudorhiza………A. japonica

     
  56. 28.

    Volval remnants on the pileus verrucose to pyramidal; stipe base distinctly radicating with long pesudorhiza………A. miculifera

     
  57. 29.

    Pileus gray to gray-brown; volval remnants on pileus usually absent; volval remnants on stipe base limbate and composed of scarce inflated cells……….30

     
  58. 29.

    Pileus white; volval remnants on pileus often retained as white patches; volval remnants on stipe base limbate and composed of abundant inflated cells………A. oberwinklerana

     
  59. 30.

    Lamellae white lacking pinkish tinge; basidiospores relatively smaller, with width mostly less than 9 μm and length less than 6 μm……….31

     
  60. 30.

    Lamellae dirty white often with pinkish tinge; basidiospores relatively larger, with width mostly more than 9 μm and length more than 6 μm, 9.0–11.0 × 6.0–8.0 μm, Q = 1.32–1.57, Qm = 1.45 ± 0.09………A. roseolifolia

     
  61. 31.

    Annulus apical to subapical, large and fragile……….32

     
  62. 31.

    Annulus median, small and non-fragile……….34

     
  63. 32.

    Basidioma relatively larger with pileus ca. 5–15 cm in diam.; basal bulb ventricose to clavate, often non-globose; basidiospores broadly ellipsoid to ellipsoid……….33

     
  64. 32.

    Basidioma relatively smaller with pileus ca. 4–6 cm in diam.; basal bulb subglobose; basidiospores globose to broadly ellipsoid, 5.5–7.5 × 5.0–6.0 μm, Q = 1.1–1.36, Qm = 1.23 ± 0.07………A. modesta

     
  65. 33.

    Basidiospores relatively longer, 7.0–9.0 × 4.5–6.0 μm, Q = 1.27–1.7, Qm = 1.49 ± 0.13; volval remnants on stipe base with fairly abundant inflated cells, globose, subglobose, ellipsoid to clavate………A. pseudoporphyria

     
  66. 33.

    Basidiospores relatively shorter, 6.0–8.0 × 4.5–6.5 μm, Q = 1.16–1.57, Qm = 1.37 ± 0.09; volval remnants on stipe base with scarce inflated cells, mostly fusiform, ellipsoid, elongate to clavate, rarely subglobose………A. caojizong

     
  67. 34.

    Basidioma relatively robust; annulus thick and persistent on stipe surface; volva on stipe base thick and gray with inner part composed of very abundant inflated cells, globose, subglobose, fusiform, ellipsoid, elongate to clavate………A. manginiana

     
  68. 34.

    Basidioma relatively slender; annulus thin and attached on stipe surface; volva on stipe base thin and white composed of fairly abundant inflated cells, mostly ellipsoid, elongate to clavate, rarely subglobose………A. pseudomanginiana

     

100. Amanita alboflavescens Hongo, Mem. Fac. Liberal Arts Shiga Univ. Nat. Sci. 20: 50 (1970).

Pileus 4–9 cm in diam., white (1A1) to dirty white (2B1), with yellow (2A2–5) color change when injured; volval remnants on pileus floccose, conical to patchy, dirty white (2B1) to grayish (1B2–3); margin non-striate, appendiculate; trama white (1A1), with yellow (2A2–5) color change when injured. Lamellae white (1A1) to cream (1A2); lamellulae attenuate. Stipe 6–9 cm long × 0.5–1.5 cm diam., white (1A1), covered with dirty white (2B1) squamules; context white (1A1), with yellow (2A2–5) color change when injured; basal bulb subfusiform, 1.5–3 cm diam., with upper part covered with patchy volval remnants. Annulus white (1A1), with yellow (2A2–5) color change when injured, fragile and fugacious.

Basidia 35–40 × 9–12 μm, clavate, 4-spored. Basidiospores [83/4/4] (7.5–) 8.0–11.0 (–12.5) × 4.5–5.5 (–7.0) μm, Q = (1.28–) 1.45–2.0 (–2.3), Qm = 1.78 ± 0.2, ellipsoid to elongate, amyloid. Volval remnants on pileus composed of irregularly arranged elements: filamentous hyphae scarce to fairly abundant; inflated cells very abundant. Volval remnants on stipe base similar to structure of volval remnants on pileus. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China (Yang 1997, 2005, 2015; this study), Japan (Hongo 1970; Imazeki and Hongo 1987; Yang and Doi 1999; Imazeki et al. 2011), Republic of Korea (Kim et al. 2013b) and Thailand (Sanmee et al. 2008).

Specimens examined: CHINA. YUNNAN PROVINCE: Chuxiong, altitude 2000 m, 24 August 1983, Wen-Kang Zhen 8349 (HKAS 12053); Fuming County, in a forest dominated by Pinus yunnanensis, altitude ca. 2050 m, 27 July 1999, Ju-Zong Li and Mu Zang 12978 (HKAS 34017); Xiaguan, Shaba, in a forest dominated by Pinus yunnanensis, altitude 2050 m, 18 September 2012, Gang Wu 1063 (HKAS 77235). JAPAN. HONSHU, Shiga-ken, Ôtsu-shi, Kamidanakami-Dô, in a forest with Quercus acutissima and Q. glauca, 18 September 1969, Hongo Herb. 4014 (HOLOTYPE, TNS-F-237274).

Commentary: Amanita alboflavescens was described from Japan by Hongo (1970), and subsequently reported from China, the Republic of Korea and Thailand (Yang 1997, 2005, 2015; Sanmee et al. 2008; Kim et al. 2013b). We examined the holotype of A. alboflavescens and the basidiospores are [20/1/1] 8.5–10.5 (–11) × (4.5–) 5–5.5 (–6) μm, Q = (1.55–) 1.66–2.09 (–2.2), Qm = 1.87 ± 0.16, elongate to subcylindrical, amyloid.

Our current phylogenetic data do not suggest any species related to A. alboflavescens (Fig. 1b). For detailed descriptions, comparisons with similar species, line drawings and images of A. alboflavescens see Yang (1997, 2005, 2015).

101. Amanita avellaneifolia Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 37a, b, 38.
Fig. 37

Fresh basidiomata of novel species in Amanita sect. Roanokenses. ab A. avellaneifolia (HKAS 79891); c–e A. brunneostrobilipes (c TYPE, HKAS 60291, de HKAS 96787); f–g A. caojizong (HKAS 100615); h–i A. elliptica (TYPE, HKAS 79602); j–k A. luteofolia (TYPE, HKAS 81895); l–m A. minutisquama (TYPE, HKAS 100504); n–o A. pallidochlorotica (TYPE, HKAS 77280). Bars: 2 cm

Fig. 38

Microscopic features of Amanita avellaneifolia (TYPE, HKAS 80011). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825040

Etymology: avellaneifolia, from avellaneus = gray-brown, and folia = leaf, referring to gray-brown lamellae of this species.

Type: CHINA. YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a forest with Pinus kesiya var. langbianensis, altitude 2000 m, 5 August 2013, Zhu L. Yang 5731 [TYPE, HKAS 80011, as Amanita sp. 13 in Yang (2015), GenBank Acc. No.: nrLSU = MH486378, tef1-α  = MH508680, rpb2 = MH485872, β-tubulin  = MH485410].

Basidioma (Fig. 37a, b) small to medium-sized. Pileus 3–7 cm diam., applanate, often slightly depressed at center, dirty white (4A2) to cream (2B2–3,), sometimes grayish (4B1) to gray-brown (4B2–4) at center; volval remnants on pileus granular to conical, ca. 1 mm in height and width, dirty white (4A2), grayish white (1A1) to dark gray (2C1); margin non-striate, appendiculate; trama dirty white (4A2), unchanging. Lamellae free, crowded, biscuit colored (4B2–5), gray-brown (4C3–5) to pale flesh colored (4A3–4); lamellulae attenuate, plentiful. Stipe 5–8 cm long × 0.5–1 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, grayish (4B1) to brownish (4B2–3), with minute concolorous squamules; context grayish (1B1), unchanging or wounded place becoming flesh color (5A2–3), stuffed in center; basal bulb subglobose, 1.5–2.3 cm diam., dirty white (1B1), with upper part covered with verrucose, dirty grayish (2B1–2) to dirty white (1B1) volval remnants arranged in irregular concentric rings. Annulus present, apical to subapical, membranous, fragile and fugacious, white (1A1). Odor like tomato juice.

Lamellar trama bilateral. Mediostratum 20–40 μm wide, composed of abundant subfusiform to ellipsoid inflated cells (65–140 × 20–30 μm); filamentous hyphae abundant, 2–9 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subfusiform to ellipsoid inflated cells (40–110 × 10–25 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 3–7 μm wide. Subhymenium (Fig. 38a) 20–35 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 14–25 × 10–20 μm. Basidia (Fig. 38a) 45–55 × 9–13 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa clamped. Basidiospores (Fig. 38b) [210/6/2] (6.5–) 8.0–10.0 (–11.5) × (6.0–) 6.5–8.5 (–10.0) μm, Q = (1.0–) 1.07–1.35 (–1.57), Qm = 1.2 ± 0.08, broadly ellipsoid, rarely subglobose or ellipsoid, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (13–52 × 12–25 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–5 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 130–250 μm thick; upper layer (88–150 μm thick) strongly gelatinized, composed of subradially to somewhat interwoven, thin-walled, yellowish brown, filamentous hyphae 4–11 (–17) μm wide; lower layer (50–150 μm thick) composed of radially and compactly arranged, yellowish brown, filamentous hyphae 6–11 μm wide; vascular hyphae scarce. Volval remnants on pileus (Fig. 38c) composed of vertically to somewhat irregularly arranged elements: filamentous hyphae scarce, 3–7 μm wide, colorless or yellowish brown, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to nearly dominant, subglobose, subfusiform to ellipsoid, 20–70 × 10–45 μm, colorless to subcolorless at upper part, becoming yellowish brown to brown towards the pileipellis, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base similar to structure of volval remnants on pileus, but with more filamentous hyphae toward the base of the verrucose squamules. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 130–400 × 20–50 μm; filamentous hyphae scattered to abundant, 4–15 μm wide; vascular hyphae scarce. Annulus composed of subradially arranged elements: inflated cells very abundant to nearly dominant, subglobose, subfusiform to ellipsoid, 20–70 × 10–25 μm, colorless or yellowish, thin-walled; filamentous hyphae scarce, 2–8 μm wide, colorless or yellowish, thin-walled; vascular hyphae scarce. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in subtropical forests under trees of Pinus kesiya var. langbianensis; basidioma occurring in summer and autumn.

Distribution: Known from southwestern China.

Additional specimen examined: CHINA. YUNNAN PROVINCE: Jingdong County, Ailaoshan, in a forest of Pinus kesiya var. langbianensis, altitude 1950 m, 24 July 2013, Xiao-Bin Liu 190 (HKAS 79891).

Commentary: Amanita avellaneifolia is characterized by its dirty white to cream pileus covered with small granular to conical, dirty white to grayish white squamules, biscuit to pale flesh-colored lamellae, broadly ellipsoid basidiospores (8.0–10.0 × 6.5–8.5 μm) and the common presence of clamps.

Our multi-locus phylogenetic analysis (Fig. 1b) indicates that A. avellaneifolia is closely related to A. rubiginosa Qing Cai et al. For comparisons between the two species see the commentary on A. rubiginosa. Most species in A. sect. Roanokenses have white, yellowish to yellow lamellae, while A. avellaneifolia can be easily distinguished by its biscuit to pale flesh-colored lamellae. Amanita hongoi Bas, described from Japan, is comparable to A. avellaneifolia on account of its similarly colored basidioma. However, A. hongoi has white to yellowish lamellae, a white, dirty white to brownish stipe densely covered with concentrically arranged, minute, concolorous squamules, and clamps are absent in all parts of its basidioma (Bas 1969; Imazeki and Hongo 1987; Yang and Doi 1999; Yang 2015).

102. Amanita brunneostrobilipes Zhu L. Yang, Yang-Yang Cui & Qing Cai, sp. nov.

Figures 37c–e, 39.
Fig. 39

Microscopic features of Amanita brunneostrobilipes (TYPE, HKAS 60291). a Hymenium and subhymenium; b basidiospores; c longitudinal section of volval remnants on pileus. Bars: ab = 10 μm, c = 40 μm

MycoBank: MB 825041

Etymology: brunneostrobilipes, from brunneus =brown, strobilinus = cone-like, and -pes = footed, referring to cone-shaped, brown basal bulb of this species.

Type: CHINA. HAINAN PROVINCE: Qiongzhong County, Limushan, in a tropical to subtropical broad-leaved forest, altitude 800 m, 4 August 2010, Zai-Wei Ge 2615 [TYPE, HKAS 60291, as Amanita sp. 12 in Yang (2015), GenBank Acc. No.: ITS = MH508281, nrLSU = MH486415, tef1-α  = MH508703].

Basidioma (Fig. 37c–e) medium-sized to large. Pileus 5–13 cm diam., plano-convex to applanate, lacking an umbo or depression at center, white (1A1) to dirty white (4A1–2); volval remnants on pileus as large, yellow-brown (4B2–3) to brown (4C3–5, 4D3–5) pyramids at center, 2–6 mm high, 1–4 mm wide, but becoming minute, white (1A1), dirty white (4A1–2) or yellowish (1B2–4) cones to powders towards margin, less than 2 mm high; margin non-striate, appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream-colored (2A2); lamellulae attenuate, plentiful. Stipe 9–17 cm long × 0.8–2 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1) and densely covered with white (1A1), yellowish (1B2–4) to pinkish (7A2) pulverulent squamules; context white to pale brownish (3B2), stuffed in center; basal bulb fusiform, strobiliform to long clavate, radicating, 4–10 cm long, 2–3 cm diam., pale yellowish brown (1B3–4) to yellow-brown (3D2–4), densely covered with recurved, concolorous squamules arranged in concentric rings. Annulus present, apical, white (1A1) to cream (2A2), fragile and fugacious. Odor of lime.

Lamellar trama bilateral. Mediostratum 40–55 μm wide, composed of abundant subfusiform, ellipsoid to clavate inflated cells (50–80 × 10–35 μm); filamentous hyphae abundant, 3–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant subfusiform to ellipsoid inflated cells (30–40 × 8–15 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 2–4 μm wide. Subhymenium (Fig. 39a) 30–55 μm thick, with 2–3 layers of subglobose to ellipsoid or irregular cells, 10–25 × 10–20 μm. Basidia (Fig. 39a) 40–50 × 8–12 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 39b) [180/14/9] (7.0–) 8.0–11.0 (–13.0) × (4.5–) 5.0–7.0 (–8.0) μm, Q = (1.29–) 1.33–1.72 (–1.89), Qm = 1.52 ± 0.11, ellipsoid to elongate, rarely broadly ellipsoid, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (25–40 × 15–30 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 3–5 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 150–200 μm thick, lacking differentiation between upper and lower layer, non-gelatinized or with upper layer slightly gelatinized, composed of radially arranged, thin-walled, subcolorless to yellowish, filamentous hyphae 3–10 (–12) μm wide, vascular hyphae scarce. Volval remnants on pileus (Fig. 39c) composed of more or less vertically to irregularly arranged elements: filamentous hyphae scarce, 2–5 μm wide, colorless to brownish, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells very abundant to nearly dominant, globose, subglobose, subfusiform to ellipsoid, 15–65 × 15–60 μm, colorless to brownish, thin-walled, terminal or in chains of 2–3; vascular hyphae scarce. Volval remnants on stipe base similar to the structure of volval remnants on pileus, but with more abundant filamentous hyphae and fairly abundant vascular hyphae. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 200–400 × 25–50 μm; filamentous hyphae scattered to abundant, 3–15 μm wide; vascular hyphae scarce. Annulus composed of subradially arranged elements: inflated cells very abundant, subglobose, fusiform to ellipsoid, 15–40 × 15–25 μm, colorless, thin-walled; filamentous hyphae abundant, 2–7 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in tropical to subtropical broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern China.

Additional specimens examined: CHINA. HAINAN PROVINCE: Qiongzhong County, Limushan, in a broad-leaved forest with Fagaceae, altitude 840 m, 21 June 2016, Yang-Yang Cui 286 (HKAS 96785); same location, in a broad-leaved forest with Fagaceae, altitude 840 m, 21 June 2016, Yang-Yang Cui 287 (HKAS 96786); same location, in a broad-leaved forest with Fagaceae, altitude 840 m, 21 June 2016, Yang-Yang Cui 288 (HKAS 96787); same location, in a broad-leaved forest with Fagaceae, altitude 840 m, 21 June 2016, Yang-Yang Cui 289 (HKAS 98788); same location, in a broad-leaved forest with Fagaceae, altitude 840 m, 26 June 2016, Yang-Yang Cui 315 (HKAS 96814); same location, in a broad-leaved forest with Fagaceae, altitude 850 m, 21 June 2016, Kuan Zhao 857 (HKAS 101441); same location, in a forest with Fagaceae, altitude 850 m, 21 June 2016, Gang Wu 1544 (HKAS 99660); same location, in a tropical broad-leaved forest, altitude 650 m, 6 May 2009, Nian-Kai Zeng 123 (HKAS 100514).

Commentary: Amanita brunneostrobilipes is characterized by its dirty white pileus densely covered with large, brown pyramids at the center, and minute, white to brownish cones to powders at the margin, a strobiliform to long-clavate stipe base covered with brown, recurved squamules arranged in concentric rings, and the absence of clamps. It is associated with broad-leaved forests.

Phylogenetically, A. brunneostrobilipes clustered as a distinct clade in A. sect. Roanokenses, away from other species (Fig. 1b). Amanita brunneostrobilipes is similar to A. timida Corner & Bas, a species described from Singapore, in the color of the basidioma and recurved squamules on the stipe base. However, the latter can be distinguished by its white pileus with a pinkish tinge, broad ellipsoid basidiospores (7.0–9.5 × 5.5–7.0 μm) and the common presence of clamps (Corner and Bas 1962; Bas 1969; Yang 2015).

Amanita brunneostrobilipes is comparable to A. strobilipes Thongbai et al. Bin in that they share a strobiliform basal bulb. However, A. strobilipes, described from Thailand, differs from A. brunneostrobilipes by its darker colored (dark gray) cones to pyramids on the pileus, a more appendiculate pileus margin and a non-radicating basal bulb (Li et al. 2016).

103. Amanita caojizong Zhu L. Yang, Yang-Yang Cui and Qing Cai, sp. nov. [Amanita manginiana sensu W. F. Chiu, Sci. Rept. Natl. Tsing Hua Univ., Ser. B, Biol. & Psychol. Sci. 3 (3): 166 (1948)]

Figures 37f, g, 40.
Fig. 40

Microscopic features of Amanita caojizong (TYPE, HKAS 79673). a Hymenium and subhymenium; b basidiospores; c longitudinal section of outer part of volval remnants on stipe base; d longitudinal section of inner part of volval remnants on stipe base;. Bars: ab = 10 μm, cd = 40 μm

MycoBank: MB 825042

Etymology: caojizong is named because it is commonly called “caojizong” in mushroom markets in Yunnan Province, China.

Type: CHINA. YUNNAN PROVINCE: Kunming, Qiongzhusi, in a forest with Pinus yunnanensis and Fagaceae, altitude 2000 m, 28 July 2013, Zhu L. Yang 5712 (TYPE, HKAS 79673, GenBank Acc. No.: ITS = MH508291, nrLSU = MH486429, tef1-α  = MH508714, rpb2 = MH485908).

Basidioma (Fig. 37f, g) medium-sized to large, sometimes very large. Pileus 5–15 (–20) cm in diam., plano-convex to applanate, sometimes umbonate at center, brownish gray (1B2–4), brown (1C3–5), gray (1D1–2) to dark gray (2D1–3), with innate, dark gray (2D1–3), radiating fibrils; volval remnants on pileus absent, or sometimes with retained, white (1A1) patches; margin non-striate, appendiculate; trama white (1A1), unchanging. Lamellae free, crowded, white (1A1) to cream (2A2); lamellulae attenuate. Stipe 8–15 cm long × 0.5–3 cm diam., subcylindric or slightly tapering upwards, with apex slightly expanded, white (1A1), covered with fibrous to pulverulent, white (1A1) squamules; context white (1A1), stuffed to soft in center; basal bulb fusiform to clavate, 2–3.5 cm diam.; volval remnants on stipe base limbate, membranous, with free limb up to 4 cm in height, outer surface white (1A1) to dirty white (1B2), inner surface white (1A1). Annulus apical to subapical, white, large, fragile and fugacious. Odor indistinct.

Lamellar trama bilateral. Mediostratum 25–45 μm wide, composed of abundant subfusiform, ellipsoid to elongate inflated cells (45–85 × 15–25 μm); filamentous hyphae abundant, 3–8 μm wide; vascular hyphae scarce. Lateral stratum composed of abundant elongate to clavate inflated cells (40–80 × 10–20 μm), diverging at an angle of ca. 30° to 45° to mediostratum; filamentous hyphae abundant and 3–7 μm wide. Subhymenium (Fig. 40a) 20–40 μm thick, with 2–3 layers of subglobose, ovoid to ellipsoid or irregular cells, 8–30 × 6–15 μm. Basidia (Fig. 40a) 25–45 × 6–11 μm, clavate, 4-spored; sterigmata 3–5 μm long; basal septa lacking clamps. Basidiospores (Fig. 40b) [200/25/20] 6.0–8.0 (–9.0) × 4.5–6.5 μm, Q = 1.16–1.57 (–1.7), Qm = 1.37 ± 0.09, broadly ellipsoid to ellipsoid, amyloid, colorless, thin-walled, smooth; apiculus small. Lamellar edge appearing as a sterile strip, composed of subglobose to ellipsoid or sphaeropedunculate inflated cells (10–30 × 8–20 μm), single and terminal or in chains of 2–3, thin-walled, colorless; filamentous hyphae abundant, 2–8 μm wide, irregularly arranged or ± running parallel to lamellar edge. Pileipellis 100–200 μm thick; upper layer (30–80 μm thick) strongly gelatinized, composed of radially, thin-walled, colorless or brownish, filamentous hyphae 2–6 μm wide; lower layer (30–120 μm thick) composed of radially and compactly arranged, filamentous hyphae 2–8 (–15) μm wide, yellowish to brownish; vascular hyphae scarce. Volval remnants on pileus dominantly composed of irregularly arranged filamentous hyphae 2–8 (–15) μm wide, mixed with elongate, fusiform to broadly clavate inflated cells (40–110 × 12–50 μm), often interjacent, occasionally terminal; outer surface non- or slightly gelatinized; inner surface strongly gelatinized. Interior of volval limb on stipe base (Fig. 40c, d) composed of longitudinally arranged elements: filamentous hyphae very abundant to dominant, 2–8 μm wide, colorless, thin-walled (≤ 0.5 μm), branching, anastomosing; inflated cells scarce to scattered, subfusiform, ellipsoid, elongate to clavate, rarely subglobose, 45–90 × 13–30 (–45) μm, colorless, thin-walled, interjacent or terminal; vascular hyphae scarce. Outer surface of volval limb similar to interior part, but with more abundant filamentous hyphae; inner surface gelatinized, similar to interior part. Stipe trama composed of longitudinally arranged, long clavate, terminal cells, 200–280 × 16–35 μm; filamentous hyphae scattered to abundant, 2–8 μm wide; vascular hyphae scarce. Annulus composed of loosely, interwoven elements: inflated cells abundant, subglobose, subfusiform to ellipsoid, 10–65 × 8–35 μm, colorless, thin-walled; filamentous hyphae scarce to fairly abundant, 2–10 μm wide, colorless, thin-walled; vascular hyphae scarce. Clamps absent in all parts of basidioma.

Habitat: Solitary to scattered in pine, broad-leaved or mixed forests with Fagaceae and Pinaceae; basidioma occurring in summer and autumn.

Distribution: Known from eastern, central, southern and southwestern China, Japan, Republic of Korea and Thailand.

Specimens examined: CHINA. ANHUI PROVINCE: Huangshan, in a broad-leaved forest, altitude unknown, 30 August 1957, Shu-Qun Deng 5178 (HMAS 20333); Jinzhai County, in a mixed forest with Fagaceae and Pinaceae, altitude 955 m, 22 July 2017, Yan-Jia Hao 1532 (HKAS 100615). FUJIAN PROVINCE: Xiamen, in a mixed forest, altitude unknown, June 1998, Xiao-Lan Mao 9411 (HMAS 74813). GUANGDONG PROVINCE: Fengkai County, Heishiding, in a subtropical forest dominated by Fagaceae, altitude 500 m, 9 May 2012, Fang Li 196 (HKAS 78421); same location, in a subtropical forest dominated by Fagaceae, altitude ca. 500 m, 10 May 2012, Fang Li 215 (HKAS 78429). GUANGXI PROVINCE: Donglan County, in a forest with Castanea mollissima, altitude unknown, 15 June 1970, Yu-Chen Zong et al. 133 (HMAS 35769). GUIZHOU PROVINCE: Fanjingshan, altitude unknown, 27 August 1982, Yu-Chen Zong and Hua-An Wen 215 (HMAS 60311). HUNAN PROVINCE: Yizhang County, in a subtropical forest, altitude and collection date unknown, Yu-Chen Zong and Xiao-Lan Mao 97 (HMAS 42199). JIANGSU PROVINCE: Nanjing, altitude unknown, 13 June 1937, Shu-Qun Deng 2729 (CUP-CH 1156). SICHUAN PROVINCE: Miyi County, altitude ca. 1890 m, 19 June 1986, Ming-Sheng Yuan 1149 (HKAS 18291). YUNNAN PROVINCE: Binchuan County, Jizushan, in a forest with Quercus and Pinus, altitude 1988 m, 11 August 2011, Li-Ping Tang 1539 (HKAS 70000); Nanjian County, Wuliangshan, in a pine forest with Pinus yunnanensis, altitude 2229 m, 28 July 2009, Li-Ping Tang 976 (HKAS 56933); Jinghong, Dadugang, in a forest with Lithocarpus, altitude 1300 m, 10 July 2014, Gang Wu 1314 (HKAS 89051); Kunming, in a forest with Lithocarpus and Castanopsis, altitude 2200 m, 8 August 2007, Zhu L. Yang 4909 (HKAS 52226); Lanping County, Hexixiang, in a pine forest with Pinus yunnanensis, altitude 2350 m, 16 August 2011, Gang Wu 727 (HKAS 75042); Xundian County, altitude 1800 m, collection date unknown, Qing Cai 675, 676 and 678 (HKAS 100501, 100502 and 100503 respectively); Yulong Naxi Autonomous County, Jiuhexiang, in a pine forest, altitude 2780 m, 20 August 2010, Qi Zhao 996 (HKAS 69717).

Commentary: This is a very well-known wild edible mushroom in Yunnan. Chiu (1948) used the name “A. manginiana Har. & Pat.” for the collections from Yunnan. However, there are some morphological discrepancies between materials from Yunnan, China and the protologue and type illustration (Hariot and Patouillard 1914; Gilbert 1941a). Therefore, Yang (1997, 2005, 2015) applied the name A. manginiana sensu W. F. Chiu to document the Chinese materials.

In our current phylogenetic study (Fig. 1b), A. manginiana sensu W. F. Chiu and A. manginiana clustered as two distinct monophyletic clades. In addition, A. manginiana sensu W. F. Chiu can be distinguished from A. manginiana by its relatively larger basidioma with pileus ca. 5–15 (–20) cm in diam., an apical to subapical, large, fragile annulus, a thin, white, limbate volva on the stipe base composed with scarce, ellipsoid, elongate to clavate, rarely subglobose inflated cells (Hariot and Patouillard 1914; Gilbert 1941a; this study). Thus, we describe A. manginiana sensu W. F. Chiu here as a novel species. The name A. caojizong is chosen because it is commonly sold in the mushroom markets of Yunnan Province, China as “caojizong”.

According to our ITS phylogenetic analysis (Fig. S12), six collections, HKAS 38460 (ITS = AY436463) from Yunnan identified as Amanita manginiana sensu W. F. Chiu by Zhang et al. (2004), FB-30951(CBM) (ITS = AB015702) from Japan identified as A. pseudoporphyria by Oda et al. (2002c), SFC20140823-10 (ITS = KT779083) from the Republic of Korea identified as A. manginiana by Cho et al. (2015), BZ-N11 (ITS = KY747457) from Thailand identified as A. cf. manginiana by Thongbai et al. (2017), and A16 (ITS = FJ441046) and A4 (ITS = FJ441034) submitted to NCBI directly as A. pseudoporphyria, should be A. caojizong.

Amanita pseudoporphyria Hongo and A. pseudomanginiana Qing Cai et al. can be confused with A. caojizong because of their gray-brown pilei and the limbate volval remnants on the stipe base. However, A. pseudoporphyria differs from A. caojizong by its relatively longer basidiospores (7.0–9.0 × 4.5–6.0 μm, Q = 1.27–1.7, Qm = 1.49 ± 0.13), and the volval remnants on the stipe base have fairly abundant, globose, subglobose to fusiform inflated cells (Hongo 1957; Imazeki and Hongo 1987; Yang 1997, 2000, 2005, 2015; Yang and Doi 1999; Yang et al. 2001; Imazeki et al. 2011). Amanita pseudomanginiana can be distinguished with A. caojizong by its relatively smaller basidioma with pileus ca. 5 cm in diam., a median annulus, a subglobose basal bulb and fairly abundant, ellipsoid, elongate to clavate inflated cells in the volval remnants on the stipe base.

Amanita griseoturcosa Oda et al., A. roseolifolia Yang-Yang Cui et al. and A. modesta Corner & Bas are also similar to A. caojizong in their appearance. However, A. griseoturcosa has a smaller basidioma with pileus 4–6 cm in diam. and larger basidiospores (8.4–12.0 × 5.6–7.6 μm, Q = 1.29–1.69, Qm = 1.50 ± 0.13) (Oda et al. 2002a; Cho et al. 2015).

Amanita roseolifolia has a relatively smaller basidioma with pileus ca. 4–7 cm in diam., dirty white lamellae often with a pinkish tinge and relatively larger basidiospores (9.0–11.0 × 6.0–8.0 μm, Q = 1.32–1.57, Qm = 1.45 ± 0.09).

Amanita modesta has a small basidioma with pileus ca. 4–6 cm in diam., a subglobose basal bulb, a thick, fleshy volva on the stipe base with abundant inflated cells and subglobose to broadly ellipsoid basidiospores (5.5–7.5 × 5.0–6.0 μm, Q = 1.1–1.36, Qm = 1.23 ± 0.07) (Corner and Bas 1962; Yang 2015; Lee 2017).

104. Amanita castanopsis Hongo, Bull. Soc. Linn. Lyon (Num. spéc.): 192 (1974).

Pileus 5–11 (–16) cm in diam., white (1A1); volval remnants on pileus conical to pyramidal, large, ca. 2–5 mm in height and 2–8 mm in width, white (1A1) to dirty white (2B1), with its apical part gray (3C1) to brownish (3D2–4) when dried; margin non-striate, appendiculate; trama white (1A1), unchanging. Lamellae white (1A1); lamellulae attenuate. Stipe 7–12 (–16) cm long × 1–2.5 cm diam., white (1A1); context white (1A1), unchanging; basal bulb napiform to somewhat radicating, 2–4 cm diam., with upper part covered with verrucose to subpyramidal, white (1A1) volval remnants, often arranged in incomplete belts. Annulus fragile and fugacious.

Basidia 45–65 × 11–15 μm, clavate, 4-spored. Basidiospores [80/4/4] (8.5–) 9.0–11.0 (–12.5) × (4.5–) 5.0–6.5 μm, Q = (1.5–) 1.54–2.02 (–2.44), Qm = 1.81 ± 0.18, ellipsoid to elongate, amyloid. Volval remnants on pileus composed of more or less vertically arranged elements: filamentous hyphae fairly abundant; inflated cells abundant. Clamps present in all parts of basidioma.

Habitat: Solitary to scattered in broad-leaved forests with Fagaceae; basidioma occurring in summer and autumn.

Distribution: Known from southern and southwestern China (Yang 1997, 2005, 2015; this study) and Japan (Hongo 1974b; Imazeki and Hongo 1987; Yang and Doi 1999; Imazeki et al. 2011).

Specimens examined: CHINA. HAINAN PROVINCE: Ledong County, in a tropical forest, altitude unknown, 11 September 1988, Huan-Qiang Chen s.n. (GDGM 15726). YUNNAN PROVINCE: Baoshan, in a forest with Castanopsis and